US7902352B2 - Isolated nucleic acid duplex for reducing huntington gene expression - Google Patents

Isolated nucleic acid duplex for reducing huntington gene expression Download PDF

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US7902352B2
US7902352B2 US11/501,634 US50163406A US7902352B2 US 7902352 B2 US7902352 B2 US 7902352B2 US 50163406 A US50163406 A US 50163406A US 7902352 B2 US7902352 B2 US 7902352B2
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nucleic acid
seq
strand
expression
vector
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US20070261126A1 (en
US20100325746A9 (en
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William F. Kaemmerer
Michael D. Kaytor
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Medtronic Inc
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Priority to EP07811178A priority patent/EP2056881A4/en
Priority to JP2009523833A priority patent/JP2010500025A/ja
Priority to CNA2007800372445A priority patent/CN101557831A/zh
Priority to PCT/US2007/017638 priority patent/WO2008021136A2/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/111—Antisense spanning the whole gene, or a large part of it
    • C—CHEMISTRY; METALLURGY
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14171—Demonstrated in vivo effect

Definitions

  • the present invention relates to inhibitory nucleic acid molecules that suppress the expression of the Huntington's disease gene in primates, including rhesus monkeys ( Macaca mulatta ) and humans ( Homo sapiens ), and methods of use thereof.
  • Huntington's disease is a neurodegenerative brain disorder with a juvenile or adult onset. It slowly destroys an affected individual's ability to walk, think, talk and reason. Symptoms include changes in cognitive ability, such as impaired short-term memory and a decreased ability to concentrate; changes in mood, such as the development of mood swings, depression and irritability; and changes in coordination and physical movement such as clumsiness, involuntary movements and twitching. These symptoms gradually worsen until HD patients die, approximately 15-20 years after the onset of the disease.
  • HD is inherited as an autosomal dominant trait. This inheritance feature means that every individual who inherits a mutated (expanded) HD gene from either parent will develop the disease.
  • the HD gene encodes for a protein called “huntingtin” (also known as “htt”).
  • huntingtin also known as “htt”.
  • the exact function of huntingtin is not known.
  • the expression of a mutant, expanded huntingtin protein is known to be the cause of HD, however.
  • mRNA messenger RNA
  • Ribosomes read the sequence of the mRNA and create the protein for which it encodes. This process of new protein synthesis is known as translation. A variety of factors affect the rate and efficiency of protein translation.
  • the intrinsic stability of the mRNA itself is the intrinsic stability of the mRNA itself. If the mRNA is degraded quickly within the cell (such as before it reaches a ribosome), it is unable to serve as a template for new protein translation, thus reducing the cell's ability to create the protein for which it encoded.
  • RNA interference is, in fact, a naturally-occurring mechanism for suppressing gene expression and subsequent protein translation. RNA interference suppresses protein translation by either degrading the mRNA before it can be translated or by binding the mRNA and directly preventing its translation. This naturally-occurring mechanism of RNA interference can also be artificially induced to occur in cells.
  • RNA interference can be achieved by introducing into cells short, double-stranded nucleic acid oligonucleotides corresponding to the mRNA for the gene to be suppressed, or by introducing into cells a sequence of DNA that encodes for a short, hairpin transcript of nucleic acids that folds back upon itself and forms a short, double-stranded nucleic acid oligonucleotide following further processing in the cell.
  • This technology provides a means to suppress the expression of huntingtin in cells.
  • the suppression of huntingtin in cells can be useful in the study of HD pathogenesis. Suppressing huntingtin in a patient also could prevent or alleviate the symptoms of HD.
  • RNA interference RNA interference
  • Suppressing expression of the HD gene can reduce levels of huntingtin within cells. This suppression and reduction can be useful in the study of HD pathogenesis. This suppression and reduction also can be useful in the prevention and treatment of the symptoms of HD.
  • RNAi is mediated by double stranded RNA (“dsRNA”), short hairpin RNA (“shRNA”) or other nucleic acid molecules with similar characteristics. These nucleic acid molecules are processed or cut into smaller pieces by cellular enzymes including Dicer and Drosha.
  • the smaller fragments of the nucleic acid molecules can then be taken up by a protein complex called the RNA-induced silencing complex (“RISC complex”) that mediates degradation of mRNAs.
  • RISC complex RNA-induced silencing complex
  • the RISC complex will degrade mRNA that complementarily base pairs with the nucleic acid molecules it has taken up. In this manner, the mRNA is specifically destroyed, thus preventing the protein for which the mRNA encoded from being made.
  • RNAi RNAi RNAi RNAi RNAi .
  • nucleic acid sequences and molecules that are homologous to primate HD mRNA sequences are introduced into cells to suppress expression of huntingtin protein.
  • nucleic acid sequences and molecules specifically suppress the expression of mRNA sequences that encode for primate huntingtin protein, including rhesus monkey and human huntingtin. Suppressing expression of this protein can be useful in the study of HD pathogenesis. This suppression also could be useful in the prevention and/or treatment of HD.
  • the invention includes a nucleic acid molecule comprising a first strand and a second strand wherein the first strand comprises a nucleotide sequence and wherein the second strand comprises the reverse complement of said first strand and wherein the nucleic acid molecule suppresses the expression of both Macaca mulatta and Homo sapiens mRNA sequences that encode for huntingtin.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 1 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 2 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 3 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 4 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 5 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 6 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 7 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 8 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 9 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 10 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 11 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 12 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 13 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 14 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 19 contiguous nucleotides encoded by SEQ ID NO: 15 and wherein the second strand is complementary to at least 15 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 9 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 10 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 11 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 12 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 13 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 14 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the invention in another embodiment, includes an isolated nucleic acid duplex comprising a first strand of nucleic acid and a second strand of nucleic acid, wherein the first strand comprises at least 27 contiguous nucleotides encoded by SEQ ID NO: 15 and wherein the second strand is complementary to at least 23 contiguous nucleotides of the first strand.
  • the nucleic acid duplex is between 19 and 30 base pairs in length.
  • the first and/or second strand of the nucleic acid duplex comprises an overhang region. In another embodiment of the present invention, the first and/or second strand of the nucleic acid duplex comprises a 3′ overhang region, a 5′ overhang region, or both 3′ and 5′ overhang regions. In another embodiment of the present invention, the first and/or second strand of the nucleic acid duplex comprises an overhang region that is from approximately 1 to approximately 10 nucleotides in length.
  • the first and second strand of the nucleic acid duplex are operably linked by means of a nucleic acid loop strand that forms a hairpin structure comprising a duplex structure and a loop structure.
  • the first and second strand of the nucleic acid duplex are operably linked by means of a nucleic acid loop that contains from 4 to 10 nucleotides.
  • the invention includes an expression cassette.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 1.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 2.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 3.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 4.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 5.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 6.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 7.
  • the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 8. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 9. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 10. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 11. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 12. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 13. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 14. In another embodiment, the expression cassette comprises a nucleic acid sequence encoding SEQ ID NO: 15.
  • the expression cassette also comprises a promoter. In another embodiment of the present invention, the expression cassette comprises a regulatable promoter. In another embodiment of the present invention, the expression cassette comprises a constitutive promoter. In another embodiment of the present invention, the expression cassette comprises a promoter that is a cytomegalovirus (“CMV”) promoter. In another embodiment of the present invention, the expression cassette comprises a promoter that is a Rous sarcoma virus (“RSV”) promoter. In another embodiment of the present invention, the expression cassette comprises a promoter utilized by RNA polymerase II. In another embodiment of the present invention, the expression cassette comprises a promoter utilized by RNA polymerase III.
  • CMV cytomegalovirus
  • RSV Rous sarcoma virus
  • the expression cassette comprises a polyadenylation signal. In another embodiment of the present invention, the expression cassette comprises a polyadenylation signal that is a synthetic minimal polyadenylation signal. In another embodiment of the present invention, the expression cassette comprises a marker gene.
  • the invention includes a vector comprising one or more of the expression cassettes previously described.
  • the vector comprises a first and a second expression cassette.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 1 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 1.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 2 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 2.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 4 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 4.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 5 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 5.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 6 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 6.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 7 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 7.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 9 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 9.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 10 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 10.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 11 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 11.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 12 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 12.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 14 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 14.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 15 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 15 contiguous nucleotides of SEQ ID. NO. 15.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 9 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 23 contiguous nucleotides of SEQ ID. NO. 9.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 10 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 23 contiguous nucleotides of SEQ ID. NO. 10.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 12 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 23 contiguous nucleotides of SEQ ID. NO. 12.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 13 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 23 contiguous nucleotides of SEQ ID. NO. 13.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 14 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 23 contiguous nucleotides of SEQ ID. NO. 14.
  • the first expression cassette of the vector encodes a nucleotide sequence encoding for SEQ ID NO. 15 and the second expression cassette of the vector encodes for a nucleotide sequence that is complementary to at least 23 contiguous nucleotides of SEQ ID. NO. 15.
  • the vector is a viral vector. In another embodiment of the present invention, the vector is an adenoviral virus vector. In another embodiment of the present invention, the vector is a lentiviral virus vector. In another embodiment of the present invention, the vector is an adeno-associated viral (AAV) virus vector. In another embodiment of the present invention, the vector is a poliovirus vector. In another embodiment of the present invention, the vector is a herpes simplex virus vector. In another embodiment of the present invention, the vector is a feline immunodeficiency virus vector. In another embodiment of the present invention, the vector is a murine Maloney-based viral vector.
  • AAV adeno-associated viral
  • the vector comprises a promoter. In another embodiment of the present invention, the vector comprises an inducible promoter.
  • the invention includes a cell comprising a previously-described expression cassette.
  • the cell is a mammalian cell.
  • Another embodiment of the present invention includes a non-human mammal comprising a previously-described expression cassette.
  • the embodiments of the present invention also include methods.
  • One method of the present invention includes a method of suppressing the accumulation of huntingtin in a cell comprising introducing a previously-described nucleic acid duplex into the cell in an amount sufficient to suppress accumulation of huntingtin in the cell.
  • accumulation of huntingtin is suppressed by at least 10%.
  • Another method of the present invention includes a method of preventing cytotoxic effects of mutant huntingtin in a cell comprising introducing a previously-described nucleic acid duplex into the cell in an amount sufficient to suppress accumulation of the mutant huntingtin so that the nucleic acid duplex prevents cytotoxic effects of mutant huntingtin in the cell.
  • Another method of the present invention includes a method of inhibiting expression of a huntingtin gene in a cell comprising introducing a previously-described nucleic acid duplex into the cell in an amount sufficient to inhibit expression of huntingtin so that the nucleic acid duplex inhibits expression of huntingtin.
  • Another method of the present invention includes a method of inhibiting expression of huntingtin in Macaca mulatta and Homo sapiens comprising providing a Macaca mulatta or Homo sapiens containing a neuronal cell that contains and expresses the huntingtin gene and is susceptible to nucleic acid interference and contacting the Macaca mulatta or Homo sapiens with a previously-described nucleic acid duplex thereby inhibiting expression of the huntingtin gene.
  • expression of huntingtin is inhibited by at least 10%.
  • Another method of the present invention includes a method of preventing cytotoxic effects of Huntington's disease (“HD”) in a Macaca mulatta or Homo sapiens comprising introducing a previously-described vector into a cell in an amount sufficient to suppress accumulation of a protein associated with HD, so that the resulting nucleic acid duplex prevents the cytotoxic effects of HD.
  • HD Huntington's disease
  • Another method of the present invention includes a method of inhibiting expression of the huntingtin gene in a Macaca mulatta or Homo sapiens comprising introducing a previously-described vector into a cell in an amount sufficient to inhibit expression of the huntingtin gene so that the resulting nucleic acid duplex inhibits expression of the huntingtin protein.
  • Another method of the present invention includes a method of inhibiting expression of huntingtin in a Macaca mulatta or Homo sapiens comprising providing a Macaca mulatta or Homo sapiens containing a neuronal cell, wherein the neuronal cell contains and expresses the huntingtin gene and is susceptible to nucleic acid interference, and contacting the Macaca mulatta or Homo sapiens with a previously-described vector, thereby inhibiting expression of the huntingtin gene.
  • Additional method of the present invention includes a method of preventing cytotoxic effects of Huntington's disease (“HD”) in a Macaca mulatta or Homo sapiens comprising introducing the isolated nucleic acid duplex comprising SEQ ID. NO: 1 or SEQ. ID. NO: 4 into a cell in an amount sufficient to suppress accumulation of a protein associated with HD, and wherein the nucleic acid duplex prevents cytotoxic effects of HD.
  • HD Huntington's disease
  • Additional method of the present invention includes a method of inhibiting expression of huntingtin in a Macaca mulatta or Homo sapiens comprising introducing the isolated nucleic acid duplex comprising SEQ ID. NO: 1 or SEQ. ID. NO: 4 into a cell in an amount sufficient to inhibit expression of huntingtin wherein said nucleic acid duplex inhibits expression of the protein associated with HD.
  • the nucleic acid duplexes or vectors are administered to the intrathecal space of the spinal cord. In another embodiment of the present invention, the nucleic acid duplexes or vectors are administered to the cerebrospinal fluid in one or more of the cerebral ventricles of the brain. In another embodiment of the present invention, the nucleic acid duplexes or vectors are administered directly into the brain tissue of the cerebral cortex. In another embodiment of the methods of the present invention, the nucleic acid duplexes or vectors are administered locally to the basal ganglia. In another embodiment of the present invention, the nucleic acid duplexes or vectors are administered specifically to the caudate nucleus and the putamen.
  • FIG. 1 shows a target plasmid for nucleic acid sequence characterization in HEK293 cells.
  • FIG. 2 shows rhesus monkey. HD mRNA suppression by siNA sequences in vitro in HEK293 cells.
  • FIG. 3 shows suppression of the endogenous rhesus HD gene at two different siNA doses in 4MBR5 cells.
  • FIG. 4 shows suppression of the endogenous rhesus HD gene at five different siNA doses using both 19 and 27 nucleotide length siNAs in 4MBR5 cells.
  • FIGS. 5 and 6 show suppression of the endogenous rhesus HD gene using both 19 and 27 nucleotide length siNAs in LLC-MK2 cells.
  • FIG. 7 also shows suppression of the endogenous rhesus HD gene using different doses of both 19 and 27 nucleotide length siNAs in LLC-MK2 cells.
  • FIGS. 8A-8D show suppression of the endogenous human HD gene at 4 different siNA doses in HeLa cells.
  • FIG. 9 shows suppression of exogenous rhesus huntingtin protein expression in LLC-MK2 cells.
  • FIGS. 10 and 11 show suppression of endogenous rhesus huntingtin protein expression in LLC-MK2 cells.
  • FIG. 12 depicts a structure and construction of anti-HD ( FIG. 12A ) and control ( FIG. 12B ) shNA sequences (antisense strands disclosed in SEQ. ID. NOS. 28 and 29, respectively in order of appearance).
  • FIGS. 13A-F show additional shNA sequences used in embodiments according to the present invention (antisense strands disclosed in SEQ. ID. NOS. 30-35, respectively in order of appearance).
  • FIG. 14 shows suppression of the endogenous rhesus HD gene in LLC-MK2 cells transfected with a plasmid expressing 19 or 27 nucleotide length shNAs with EB4 or mir23 loop structures.
  • FIG. 15 shows a schematic description of a pSilencer 1.0-U6 plasmid that can be used in the preparation of anti-HD and control shNA sequences.
  • FIG. 16 shows an exemplary format of AAV viral constructs.
  • FIG. 17 shows a plasmid used to construct pAAV vectors and to generate AAV in accordance with the present invention.
  • FIGS. 18A-B show additional shNA sequences used in embodiments according to the present invention (antisense strands disclosed in SEQ. ID. NO. 36 and 37, respectively, in order of appearance).
  • FIG. 19 shows suppression of exogenous rhesus HD by pAAV plasmids.
  • FIG. 20 shows suppression of endogenous human HD in HeLa cells and HEK293T cells by recombinant adeno-associated virus (AAV) expressing anti-HD shNA.
  • AAV adeno-associated virus
  • FIGS. 21 and 22 show in vivo suppression of rhesus HD gene expression by recombinant adeno-associated virus (AAV) expressing anti-HD shNA.
  • AAV adeno-associated virus
  • SEQ ID NO: X refers to, in one embodiment, each number's sequence as defined in Table 1 (identified sequence). SEQ ID NO: X must also be read to encompass sequences that would hybridize with the complementary strand of a sequence set forth in SEQ ID NOS: 1-15 and reduce the particular SEQ ID NO:'s target mRNA in a cell type selected from, without limitation, HEK293 cells, 4MBR5 cells, LLC-MK2 cells, HeLA cells or any other Macaca mulatta or Homo sapien cell type.
  • claimed sequences can include at least 99% sequence homology with the identified sequence; at least 98% sequence homology with the identified sequence; at least 95% sequence homology with the identified sequence; at least 90% sequence homology with the identified sequence; or at least 85% sequence homology with the identified sequence.
  • nucleic acid or “nucleic acid molecules” refer to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, composed of monomers (nucleotides) containing a sugar, phosphate and a base that is either a purine or pyrimidine. Unless specifically limited, the term encompasses nucleic acids containing known analogs of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also encompasses conservatively modified variants thereof.
  • the nucleic acid molecules of the present invention can include any type of nucleic acid molecule capable of mediating RNA interference, such as, without limitation, short interfering nucleic acid (siNA), short hairpin nucleic acid (shNA), short interfering RNA (siRNA), short hairpin RNA (shRNA), micro-RNA (miRNA), and double-stranded RNA (dsRNA).
  • the nucleic acid molecules of the present invention also include similar DNA sequences.
  • the nucleic acid and nucleic acid molecules of the present invention can contain unmodified or modified nucleotides. Modified nucleotides refer to nucleotides which contain a modification in the chemical structure of a nucleotide base, sugar and/or phosphate.
  • expression cassette means a nucleic acid sequence capable of directing expression of a particular nucleotide sequence in an appropriate host cell, with additional sequences that facilitate appropriate transcription of the nucleic acid sequence of interest.
  • the expression cassette can include a promoter operably linked to the nucleotide sequence of interest that also can be operably linked to termination signals.
  • the expression cassette also can include expression enhancers.
  • the expression cassette including the nucleotide sequence of interest can be chimeric.
  • the expression cassette also can be one that is naturally occurring but has been obtained in a recombinant form useful for heterologous expression.
  • the expression of the nucleotide sequence in the expression cassette can be under the control of a constitutive promoter or of a regulatable promoter that initiates transcription only when the host cell is exposed to some particular stimulus.
  • the promoter also can be specific to a particular tissue or organ or stage of development.
  • promoter refers to a nucleotide sequence, usually upstream (5 prime) of the nucleotide sequence of interest, which directs and/or controls expression of the nucleotide sequence of interest by providing for recognition by RNA polymerase and other factors required for proper transcription.
  • promoter includes (but is not limited to) a minimal promoter that is a short DNA sequence comprised of a TATA-box and other sequences that serve to specify the site of transcription initiation, to which regulatory elements are added for control of expression.
  • promoter also refers to a nucleotide sequence that includes a minimal promoter plus regulatory elements that is capable of controlling the expression of a coding sequence or functional RNA.
  • promoter sequence consists of proximal and more distal upstream elements, the latter elements often referred to as enhancers.
  • the term “enhancer” refers to a DNA sequence that can stimulate promoter activity and can be an innate element of the promoter or a heterologous element inserted to enhance the level or tissue specificity of a promoter. Enhancers are capable of operating in both orientations (normal or flipped), and are capable of functioning even when moved either upstream or downstream of the promoter. Both enhancers and other upstream promoter elements bind sequence-specific DNA-binding proteins that mediate their effects. Promoters can be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even be comprised of synthetic DNA segments.
  • a promoter also can contain DNA sequences that are involved in the binding of protein factors that control the effectiveness of transcription initiation in response to physiological or developmental conditions.
  • Specific promoters used in accordance with the present invention can include, for example and without limitation pol II promoters (including, without limitation cytomegalovirus (“CMV”) promoters, chicken ⁇ -actin (“CBA”) promoters, Rous sarcoma virus (“RSV”) promoters and neuron-specific enolase (“NSE”) promoters).
  • CMV cytomegalovirus
  • CBA chicken ⁇ -actin
  • RSV Rous sarcoma virus
  • NSE neuron-specific enolase
  • promoters used in accordance with the present invention can include, for example and without limitation, pol III promoters (including, without limitation, human H1 and human or murine U6 promoters, as well as H1 and U6 promoters engineered to be expressed in a regulated way such as described in United States Patent Application Number 2005/0064489).
  • pol III promoters including, without limitation, human H1 and human or murine U6 promoters, as well as H1 and U6 promoters engineered to be expressed in a regulated way such as described in United States Patent Application Number 2005/0064489).
  • vector is defined to include any virus, as well as any plasmid, cosmid, phage, binary vector or segment of nucleic acid (DNA or RNA) in double or single stranded linear or circular form that may or may not be self transmissible or mobilizable, and that can transform eukaryotic host cells either by integration into the cellular genome or by existing extrachromosomally (e.g., autonomous replicating plasmid with an origin of replication).
  • the gene involved in Huntington's disease (“HD”) (IT15) is located on chromosome 4 at the end of the short arm. This gene encodes for the protein huntingtin (also known as “htt”).
  • the mutation in the HD gene responsible for HD is an unstable expanded CAG trinucleotide repeat within the coding region of the gene. This mutation results in a huntingtin protein with an expanded glutamine sequence. While the normal and abnormal functions of huntingtin are not known, the presence of the mutated form of huntingtin has been correlated with the occurrence of HD symptoms. Further, the abnormal huntingtin protein appears to accumulate abnormally in neuronal nuclei. Thus, blocking the expression of huntingtin provides a useful mechanism to study HD pathogenesis and can also provide a potential treatment for HD.
  • nucleic acid sequences of their study is homologous with both rhesus and human HD genes
  • nucleic acid sequences of the present invention are known to be homologous with both rhesus ( Macaca mulatta ) and human ( Homo sapiens ) HD genes.
  • one important benefit of the present invention is that the same nucleic acid sequences can be studied and characterized for toxicological and pharmacological properties and safety in the non-human primate Macaca mulatta and then, without alteration, can also be used for therapeutic benefit in Homo sapiens.
  • RNA interference RNA interference
  • a PCR strategy was used to clone and assemble 3437 bp of the 5′ portion of the rhesus HD gene.
  • a series of PCR primers predicted to anneal to the rhesus HD cDNA were designed based on evolutionarily conserved sequences identified in an alignment of the human (NM — 002111) and pig (AB016793) cDNA sequences.
  • primers were used to amplify partially overlapping portions of the rhesus HD gene from first strand cDNA prepared from rhesus brain and kidney tissues (BioChain Institute, Inc; C1534035 and C1534142 respectively) using high fidelity DNA polymerases.
  • the recovered PCR products were cloned into pCR-Blunt-TOPO (Invitrogen) and sequenced using standard methods. Sequence alignments between multiple, independently derived clones were analyzed to ensure the elimination of potential PCR and sequencing errors. This sequence information was also used to design additional primers to clone gaps in the recovered HD cDNA sequence.
  • a single composite clone containing 3437 bp of the rhesus HD cDNA was assembled using PCR and conventional cloning techniques.
  • nucleic acid suppressor sequences for rhesus monkey huntingtin that correspond with portions of the human sequence (i.e., target genes were rhesus and human HD genes) were identified using online software (http://www.dharmacon.com/) and also obtained from Dharmacon® (Dharmacon Research, Inc., Boulder, Colo.). These nucleic acid sequences included those in the following Table 1:
  • sequences in the preceding table can be considered target sequences as well as sequences included in the nucleic acid molecules of the present invention.
  • SEQ ID NOS. 1-8 are 19 nucleotide base sequences.
  • SEQ ID NO. 9-15 are the same sequences extended to 27 nucleotide base sequences. The sequence identified as SEQ ID NO. 7 was not extended into a 27 nucleotide base sequence because extending this sequence resulted in a mismatch between the rhesus monkey and human HD sequences.
  • Control sequences also were chosen.
  • the control sequences included a non-sense scrambled control (SEQ ID NO. 19; TAGCGACTAAACACATCAA) and a TNF ⁇ sequence (SEQ ID NO. 16; AATCCTCCTTCGTATTATA) both purchased from Dharmacon.
  • rhesus monkey HD gene sequences were subcloned into pTracerTM-CMV2 (Invitrogen, Corp., Carlsbad, Calif.) to generate pTracer-rhuntingtin (pTRACER-rhHD).
  • the recombinant plasmid also included a GFP-Zeocin reporter gene for transfection efficiency normalization.
  • the CMV promoter directed constitutive expression of the target gene (rhesus monkey HD) while the EF1 promoter directed constitutive expression of the GFP-Zeocin reporter gene.
  • the generated recombinant plasmids were used to facilitate screening of nucleic acid sequences by co-transfection into a eukaryotic cell line.
  • HEK293 cell cultures at 60-70% confluency were co-transfected with the appropriate target plasmid (2 ⁇ g/well of a 6-well plate) and test nucleic acid sequences (100 nM) directed against rhesus monkey HD (SEQ ID NO: 1 through SEQ ID NO: 8), TNF ⁇ (as a control (SEQ ID NO. 16) or a non-sense scrambled control (SEQ ID NO. 19).
  • SEQ ID NO: 1 through SEQ ID NO: 8 test nucleic acid sequences directed against rhesus monkey HD
  • TNF ⁇ as a control (SEQ ID NO. 16)
  • SEQ ID NO. 19 a non-sense scrambled control
  • Forty-eight hours post-transfection total cellular RNA was harvested from the cells and used to make cDNA
  • FIG. 2 shows the results of the rhesus monkey HD transfection studies related to each of the test nucleic acid sequence's ability to suppress rhesus monkey HD mRNA. As measured by realtime PCR, all non-control sequences effectively suppressed expression of the HD gene when compared to mock treated control cells (set at 100% expression). Because the nucleic acid sequences evaluated in the present experiment have complete sequence identity to portions of the human HD gene. sequence, it is reasonable to expect that they also suppress the expression of the human HD gene, thus suppressing production of human huntingtin protein.
  • the cDNAs were used to analyze for rhesus HD and control gene expression using realtime PCR methods.
  • FIG. 3 all sequences directed against rhesus HD mRNA suppressed rhesus HD gene expression at both doses when compared to controls.
  • 4MBr5 cells were transfected as described above but with 100 nM, 10 nM, 1.0 nM, 0.2 nM or 0.05 nM of SEQ ID NO: 4 (19 nucleotide sequence); 100 nM, 10 nM, 1.0 nM, 0.2 nM or 0.05 nM of SEQ ID NO: 12 (corresponding 27 nucleotide sequence); 100 nM SEQ ID NO: 19 (scrambled) or 100 nM SEQ ID NO: 16 (off-target TNF ⁇ control). As shown in FIG.
  • LLC-MK2 cells (a rhesus kidney cell line also obtained from ATCC) at 70-80% confluency were transfected with test nucleic acid sequences (SEQ ID NO: 4 and SEQ ID NO: 12) at one of two doses, 10 nM or 0.2 nM as well as controls (SEQ ID NO: 19 and SEQ ID NO: 16) at 100 nM.
  • the cDNAs were used to analyze for rhesus HD and control gene expression using realtime PCR methods.
  • FIG. 5 all sequences directed against rhesus HD mRNA suppressed rhesus HD gene expression at both doses when compared to controls.
  • LLC-MK2 cells were transfected with 100 nM, 10 nM, 1 nM, 0.1 nM or 0.01 nM SEQ ID NO: 1 (19 nucleotide sequence); 10 nM, 1 nM, 0.1 nM or 0.01 nM SEQ ID NO: 9 (corresponding 27 nucleotide sequence); or control SEQ ID NOS: 19 or 16 (100 nM). As shown in FIG.
  • a subset of the identified siNA sequences was tested for the chosen sequence's ability to suppress expression of the endogenous human HD gene.
  • HeLa cells obtained from ATCC at 70-80% confluency were transfected with test nucleic acid sequences (SEQ ID NO: 1; SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; or SEQ ID NO: 7) or control (SEQ ID NO: 19) at 100 nM.
  • test nucleic acid sequences SEQ ID NO: 1; SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; or SEQ ID NO: 7
  • control SEQ ID NO: 19
  • Protein was harvested 48 hours post-transfection and was analyzed using western blot analysis. In this analysis, 10 ⁇ g of total protein was loaded per well. Rhesus huntingtin protein expression was measured using the Chemicon® International (Temecula, Calif.) huntingtin antibody MAB2168 and ⁇ -actin was measured with abcam® (Cambridge, Mass.) antibody 20272-100. To quantify huntingtin protein expression the intensities of the bands for huntingtin and ⁇ -actin on the autoradiogram of the western blot were measured by densitometry. Huntingtin protein expression was normalized to ⁇ -actin expression. As can be seen in FIG.
  • both SEQ ID NO: 1 and SEQ ID NO: 4 were effective to suppress exogenous huntingtin protein expression when compared to controls transfected with the pTRACER-rhHD plasmid and SEQ ID NO: 19 or the pTRACER-rhHD plasmid alone.
  • LLC-MK2 cells were transfected with 100 nM of either SEQ ID NO: 1, SEQ ID NO: 4 or SEQ ID NO: 19.
  • An additional control consisted of extracts from untransfected cells. Protein was harvested and analyzed 48 hours post-transfection with 10 ⁇ g of total protein loaded per western blot well. Again, rhesus huntingtin protein expression was measured using the Chemicon® International (Temecula, Calif.) huntingtin antibody MAB2168 and ⁇ -actin was measured with abcam® (Cambridge, Mass.) antibody 20272-100.
  • both SEQ ID NO: 1 and SEQ ID NO: 4 suppress huntingtin protein expression.
  • the level of rhesus huntingtin protein expression was normalized to ⁇ -actin expression.
  • the intensities of the bands on the autoradiogram of the western blot were measured by densitometry.
  • SEQ ID NO: 1 and SEQ ID NO: 4 both suppressed endogenous huntingtin protein expression when compared to cells transfected with control SEQ ID NO: 19 or to extract from untransfected cells.
  • FIG. 12 shows SEQ ID NO: 4 reformatted and constructed as an anti-huntingtin shNA sequence (SEQ ID NO: 17; FIG. 12A ).
  • FIG. 12 also shows a control shNA sequence (SEQ ID NO: 18; FIG. 12B ).
  • the shNA sequences contain the siNA nucleic acid sequence (boxed sequence shown in black; in FIG. 12A , an originally effective nucleic acid sequence), a nine nucleotide loop (underlined), the reverse complement of the boxed nucleic acid sequence (italicized), and a Pol III terminator sequence (TTTTTT (bolded)).
  • oligonucleotides can be used to assemble the shNAs.
  • the 1 ⁇ 3 and 2/4 oligonucleotides for each shNA can be annealed.
  • the A1 and B1 oligonucleotides include the boxed sequences.
  • the A2 and B2 oligonucleotides include the underlined nine nucleotide loop, the reverse complement of oligonucleotides A1 and B1 respectively, the bolded Pol III terminator sequence (TTTTTT) and the first G of the EcoRI site depicted in these FIGS.
  • the A3 and B3 oligonucleotides of FIGS. 12A and 12B respectively include the ApaI overhang region, the reverse complement of the boxed A1/B1 sequences, and the reverse complement of the underlined nine nucleotide loop sequence.
  • the A4 and B4 oligonucleotides include the reverse complement of the italicized portion of A2 and B2 respectively and the reverse complement (AAAAAA) of the bolded Pol III terminator sequence. Finally, the A4 and B4 oligonucleotides contain the EcoRI overhang region. ApaI and EcoRI restriction enzyme-compatible ends are included in the shNA structures for directional subcloning into a murine U6 promoter-containing shuttle vector (pSilencer1.0-U6; Ambion, Inc., Austin, Tex.; FIG. 15 ).
  • the full-length shNAs (SEQ ID NOS: 17 and 18) then can be cloned into a ApaI/EcoRI-digested pSilencer vector using a three-way ligation reaction.
  • the U6 promoter (murine or human) is used for constitutive high level expression of the nucleic acid sequences.
  • a human H1 promoter can also be used.
  • a shuttle vector containing the human U6 promoter is psiSTRIKETM hmGFP from Promega Corporation (Madison, Wis.).
  • a shuttle vector containing the human H1 promoter is pSilencer 3.0-H1 from Ambion, Inc, (Austin, Tex.).
  • the loop structure of the shNA hairpin is 4 to 10 nucleotides in length.
  • the arms of the hairpin structures are less than approximately 30 nucleotides in length.
  • the arms of the hairpin structure are between 19 and 27 nucleotides in length.
  • SEQ ID NO: 21 is a modified version of control SEQ ID NO: 18.
  • SEQ ID NO. 20 SEQ ID NO. 21
  • SEQ ID NO. 22 FIG. 13C ; modified SEQ ID NO: 17-EB4 (as will be recognized by one of ordinary skill in the art, SEQ ID NO: 22 includes SEQ ID NO: 17 with modified overhang regions for subcloning into a different vector)
  • SEQ ID NO: 23 FIG. 13D ; modified SEQ ID NO: 17-mir23
  • SEQ ID NO: 24 FIG.
  • FIG. 13E shNA-modified SEQ ID NO: 12-EB4; and SEQ ID NO: 25 ( FIG. 13F ; shNA-modified SEQ ID NO: 12-mir23).
  • FIG. 14 all test plasmids suppressed rhesus HD gene expression when compared to control plasmids.
  • the BamHI fragment ( FIG. 15 ) containing the shNA expression cassette (murine U6 promoter, shNA sequence, and Pol III terminator sequence) from each of the pSilencer shuttle vectors (huntingtin and control) is recovered, blunted with T4 DNA polymerase, and subcloned into an AAV1/2 expression vector (deprAVETM; GeneDetect.com Ltd, Bradenton, Fla.). As shown in FIG.
  • the U6 promoter (murine or human) can drive the expression of the shNA and the Woodchuck enhancer/chicken ⁇ -actin promoter can drive the expression of the enhanced green fluorescent protein (WPRE/CAG-eGFP).
  • WPRE/CAG-eGFP enhanced green fluorescent protein
  • These expression cassettes can be flanked by viral inverted terminal repeats (ITR).
  • ITR viral inverted terminal repeats
  • WPRE Woodchuck post-transcriptional regulatory element
  • BGH bovine growth hormone
  • the expression cassette additionally contains a polyadenylation signal, such as a synthetic minimal polyadenylation signal.
  • the H1 promoter can be used.
  • FIG. 17 depicts a different plasmid, the plasmid used to construct pAAV vectors used in the following experiments to generate AAV (pAAV-hrGFP, Stratagene, La. Jolla, Calif.).
  • Six different plasmids were constructed; 3 with an H1 promoter and 3 with a U6 promoter (human H1 promoter from Ambion (Austin, Tex.) and human U6 promoter from Promega (Madison, Wis.)).
  • U6-SEQ ID NO: 17 The following were subcloned into the depicted vector (1) U6-SEQ ID NO: 17; (2) U6-SEQ ID NO: 26 (SEQ ID NO: 26 is SEQ ID NO: 1 in an shRNA format; see FIG.
  • SEQ ID NO: 27 is a scrambled SEQ ID NO: 1 shNA control; see FIG. 18B ).
  • This PvuII fragment was then subcloned into the blunted MluI site of pAAV-hrGFP (Stratagene, La. Jolla, Calif.). Directionality was confirmed by restriction digest mapping and sequence analysis.
  • pAAV plasmids were co-transfected into HEK293T cells along with pTRACER-rhHD ( FIG. 1 ).
  • siRNAs SEQ ID NOS: 1 or 4
  • SEQ ID NO: 19 the non-sense scrambled control
  • pTRACER-rhHD was also transfected alone as a control.
  • RNA was collected 48 hours post transfection and the suppression of rhesus HD gene expression (on the pTRACER plasmid) was measured by realtime PCR and normalized to the amount of GFP expressed from the pTRACER-rhHD plasmid. As can be seen in FIG.
  • pAAV plasmids containing U6-SEQ ID NO: 17; U6-SEQ ID NO: 26; H1-SEQ ID NO: 17; and H1-SEQ ID NO: 26 were as effective at suppressing exogenous rhesus HD gene expression as previously tested SEQ ID NO: 1 and SEQ ID NO: 4.
  • pAAV plasmids expressing scrambled control sequences did not suppress exogenous rhesus HD gene expression.
  • AAV adeno-associated virus
  • Five separate AAV viruses were generated. These viruses were engineered to express U6-SEQ ID NO: 26; U6-SEQ ID NO: 17; H1-SEQ ID NO: 26; U6-SEQ ID NO: 18 (shNA control) or H1-SEQ ID NO: 27 (shNA control). The ability of these viruses to suppress endogenous human HD gene expression in HeLa cells and HEK293T cells was examined.
  • the cDNAs were analyzed for HD and GAPDH expression levels by realtime PCR methods. Endogenous HD expression was normalized to endogenous GAPDH expression. As shown in FIG. 20 , U6-SEQ ID NO: 26 and U6-SEQ ID NO: 17 were effective at suppressing endogenous human HD gene expression. P values were determined by comparison to the mock transduced data set. In this experiment, even though H1-SEQ ID NO: 26 did not significantly suppress endogenous human HD expression in vitro, its potential effectiveness in vivo should not be discounted especially in light of literature indicating a potential disconnect between in vitro and in vivo activity. See, for example, Wooddell et al., 334 Biochem. and Biophys. Res. Comm. 117-27 (2005).
  • the siNA sequences and molecules of the present invention can be manipulated to enhance their uptake into the RISC complex.
  • manipulating the 3 prime terminal nucleotide of the sense strand can be highly advantageous.
  • Preferential entry of the guide, or antisense, strand into RISC can be achieved by introducing 3 prime mismatches in the sense strand while maintaining perfect base pairing (of the antisense strand and the intended mRNA target) at the 5 prime terminus of the antisense strand. This maximizes entry of the antisense strand into the RISC complex, while also reducing potential off-target inhibition by the sense strand.
  • nucleic acid molecules and/or their carriers i.e. vectors
  • Some of these methods include, without limitation, calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, cationic lipid-mediated transfection, lipofection, protoplast fusion, particle bombardment, microinjection, liposome fusion, biolistics and other suitable methods found in, for example, Sambrook, et al. (Molecular Cloning: A Laboratory Manual, 2nd, ed, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals.
  • viral vectors For mammalian gene therapy, viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., rhesus monkey or human cells.
  • Other viral vectors can be derived from poxviruses, herpes simplex virus I, adenoviruses and adeno-associated viruses, and the like (see, for example, Boenisch et al., U.S. Pat. No. 5,350,674 and Wilson et al., U.S. Pat. No. 5,585,362 which are hereby incorporated by reference).
  • Embodiments of the present invention also can be delivered through the use of liposomes, polyethyleneimine, by iontophoresis, or by incorporation into other vehicles, such as hydrogels, cyclodextrins, biodegradable nanocapsules, and bioadhesive microspheres.
  • Nucleic acid molecules also can be directly delivered to cells or tissues with or without the aforementioned vehicles.
  • the nucleic acid molecules/vehicle combinations can be locally delivered by catheter and drug pump systems, delivered by direct local injection or through the use of polymers and/or drug-eluting stents.
  • the nucleic acid sequences, molecules, expression cassettes and vectors of the present invention can be used to suppress HD gene expression and resulting huntingtin formation.
  • the nucleic acid sequences, molecules, expression cassettes and vectors of the present invention also can be used in the study of HD pathogenesis.
  • the nucleic acid sequences, molecules, expression cassettes and vectors of the present invention also may be administered to prevent or treat the symptoms of HD.
  • the amount of these agents administered as well as the timing of their delivery will vary depending on various factors including, for example and without limitation, the composition chosen, the weight, physical condition, and age of the patient, and whether prevention or treatment is desired as well as other factors known to skilled practitioners.
  • Administration of the therapeutic agents can be continuous or intermittent.
  • the administration of the agents of the invention can be essentially continuous over a preselected period of time or can be in a series of spaced doses. Further, both local and systemic administration can be appropriate for use within the present invention. Such factors can be determined by the researcher or treating physician.
  • a pre-clinical study of the safety of suppressing huntingtin protein expression in the normal adult rhesus monkey brain was conducted.
  • a 6 year old female rhesus monkey weighing approximately 3.9 kg was anesthetized and bilaterally infused with an anti-HD shRNA/GFP-containing recombinant adeno-associated virus (AAV) (AAV-U6-SEQ ID NO: 17 (serotype AAV1; 2.07 ⁇ 10 12 vg/ml; lot #J0531)) by stereotactic injection.
  • AAV anti-HD shRNA/GFP-containing recombinant adeno-associated virus
  • Injections Halton syringe (100 ⁇ l) with 27 Ga needle (compression fitting)) were performed at 2 putamen sites (50 ⁇ l/site) using a single needle tract and 1 caudate site (50 ⁇ l) at a rate of 1 ⁇ l/min. Following this injection, the needle was left in place for at least 20 minutes and then withdrawn at a rate of 1 mm/min.
  • the animal was deeply anesthetized with pentobarbital (1.5 ml intramuscular; 2 ml, intravenous) and transcardially perfused with heparinized ice-cold saline (4-6 L). Following perfusion and removal, the brain was placed into a container of ice-cold saline.
  • the right hemisphere of the brain was sectioned into 14-4 mm thick coronal sections and numbered 1-14 (rostral to caudal).
  • a 14G biopsy needle was then used to collected tissue punches from various brain regions.
  • a total of 103 tissue punches was collected from the right hemisphere. Each tissue punch contained about 6 mg of tissue and was snap frozen in liquid nitrogen and stored at ⁇ 80° C. until RNA was isolated.
  • the left hemisphere of the brain was sectioned into 4 blocks (rostral to caudal).
  • Block 1 was 10 mm; blocks 2 and 3 were 16 mm; and block 4 was 20 mm.
  • Each block was embedded in OCT, snap frozen in isopentane and stored at ⁇ 80° C.
  • Ten micron sections were then collected and placed onto laser microdissection (LMD) compatible slides. Tissue on the slides was fixed in ethanol (30 seconds in 75% ethanol; 30 seconds in 95% ethanol; and 30 seconds in 100% ethanol) with an air dry between steps. Slides were then screened to identify the ones containing brain regions of interest (i.e. caudate and putamen) and data on viral spread was obtained by following the expression of the marker gene (GFP).
  • GFP positive regions were collected by LMD from the caudate and putamen.
  • a pre-clinical study of the safety of suppressing huntingtin protein expression in the normal adult rhesus monkey brain is conducted over a 12-month period.
  • six rhesus macaques ( Macaca mulatta ) are trained to perform the tasks of a commercially available computerized behavioral test battery (Monkey CANTAB, Model 80650, Lafayette Instruments, Lafayette, Ind.) that is based upon human neuropsychological tests (CANTAB, CeNeS, Cambridge, UK).
  • the performance of these six monkeys on the test battery is compared to previously published performance norms for rhesus monkeys to verify that the monkey's cognitive abilities are within normal limits. See, for example, Weed et al., Brain Research: Cognitive Brain Research, 1999, Oct. 25; 8(3) 185-201.
  • the monkeys are randomly assigned to one of two experimental groups: the first group of three monkeys receives intracranial injections of an adeno-associated virus (AAV) vector comprising an expression cassette containing an RNA polymerase III promoter driving expression of a short hairpin RNA transcript encoding for the shNA of SEQ ID NO: 17.
  • a second expression cassette in the same vector encodes for a green fluorescent protein reporter gene.
  • the second group of three monkeys receives intracranial injections of an adeno-associated virus vector comprising an expression cassette containing the same RNA polymerase III promoter driving expression of a control shNA sequence (SEQ ID NO:18).
  • a second expression cassette in this vector encodes for a green fluorescent protein reporter gene.
  • the AAV vector is administered to the caudate nucleus and putamen of each hemisphere of each monkey's brain.
  • the amount of AAV vector administered in each intracranial injection, and the surgical method and procedures for making these injections, are well-known to those skilled in the art.
  • the monkeys are repeatedly tested on a periodic basis using the Monkey CANTAB computerized testing battery for the remainder of the twelve-month study period.
  • the cognitive and behavioral performance of each monkey over time is compared to its own pre-operative performance. No statistically significant decline in cognitive or behavioral performance measures is obtained for either the monkeys treated with the control shNA vector, or the monkeys treated with the anti-rhesus-HD shNA vector. This indicates that there are no significant cognitive or behavioral consequences of suppressing the expression of huntingtin protein in the caudate and putamen of the adult primate brain.
  • Periodic neurological examinations of each monkey by skilled and qualified veterinarians unaware of the experimental group membership of each monkey also shows no neurological deficits in the monkeys.
  • the monkeys are humanely terminated, and the brain of each monkey is removed and examined for abnormalities by trained pathologists. In addition, it is studied by fluorescence microscopy, immunohistochemistry, and molecular and biochemical assays.
  • the regions of the brain in which the AAV vector effectively delivered the shNA are identified by fluorescence microscopy. This reveals expression of the green fluorescent protein reporter gene in substantial regions of the caudate nucleus and putamen in each hemisphere of each monkey's brain.
  • Immunohistochemical staining for huntingtin protein shows reduced levels of huntingtin protein in the caudate and putamen of the monkeys who received the anti-rhesus-HD shNA vector, compared to the monkeys who received the control shNA vector, indicating suppression of huntingtin protein expression by the shNA of the invention.
  • homogenized brain tissue samples from the caudate and putamen of each monkey's brain are assayed for levels of rhesus HD mRNA measured by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) assay, and for levels of rhesus huntingtin protein measured by Western blotting, using methods well-known to those skilled in the art.
  • qRT-PCR quantitative reverse transcriptase polymerase chain reaction
  • the same shNA expression cassette that is used in the pre-clinical safety tests in rhesus monkeys is engineered into an AAV vector using methods well-known to those skilled in the art.
  • This AAV vector comprises an expression cassette containing an RNA polymerase III promoter driving expression of a short hairpin RNA transcript encoding for the shNA of SEQ ID NO: 17.
  • the AAV vector optionally does not contain a second expression cassette encoding for a green fluorescent protein reporter gene, because this expression cassette is not needed for the treatment of HD in patients.
  • an AAV vector comprising an expression cassette containing SEQ ID NO: 17 is effective at suppressing the expression of the Homo sapiens HD gene and huntingtin protein, as well as the Macaca mulatta HD gene and huntingtin protein.
  • the AAV vector can be administered to the human patient's brain according to the systems and methods disclosed in U.S. Patent Application Numbers 20040162255 and 20040220132. These trials establish that suppression of huntingtin protein in the caudate and putamen of human patients afflicted with HD using the nucleic acid molecules, expression cassettes, or vectors of the present invention is effective at arresting and partially reversing the cognitive and motor deficits in patients treated after onset of these symptoms of the disease.
  • 3*10 11 viral particles comprising AAV vectors including shNAs of SEQ ID. NO: 26 or SEQ. ID. NO: 17 or a control shNA (SEQ. ID. NO: 18) under regulation of the human U6 promoter upstream of GFP sequence under control of CMV promoter were injected into Rhesus moneys as follows:
  • Huntington (HD) mRNA was quantified by qPCR using total RNA isolated from tissue punches and laser microdissected sections (LMD). Huntingtin protein was quantified by western blot analysis using total protein isolated from tissue punches.
  • the amount of HD mRNA was decreased by about 65% to 70% in the GFP-expressing areas compared to the areas not expressing GFP, as measured by qPCR of RNA isolated from LMD sections.
  • the animals were injected according to the protocol of the previous example. Histopathological analyses were conducted by analyzing GFP fluorescence, hematoxylin-eosin (H&E) staining, huntingtin immunofluorescent staining, calnexin immunofluorescent staining and protein disulfide isomerase (PDI) immuofluorescent staining.
  • H&E hematoxylin-eosin
  • PDI protein disulfide isomerase
  • the animals were injected according to the protocol of Working Example 1. Spontaneous activity and fine motor activity were also measured by EthnoVision and mMAP equipment, respectively.
  • EthnoVision is a video tracking system that can be used to measure distance traveled, body movement speed and vertical activity. Associated computer software is able to quantify each of these parameters.
  • mMAP monkey Movement Analysis Panel
  • the present invention is not limited to the particular embodiments, materials, and examples described herein, as these can vary.
  • the nucleic acid molecules of the present invention can be created in a variety of formats and lengths. Indeed, those skilled in the art will recognize that the most important attribute of the nucleic acid molecules of the present invention is their ability to enter the RISC complex and also complementarily bind to the mRNA sequences of interest to induce an RNA interference effect, resulting in reduction of the mRNA stability and/or the translation rate of these sequences.
  • the phrase “complementarily bind” as used herein, refers to the abilities of the nucleic acid molecules to form hydrogen bond(s) with mRNA sequences by either traditional Watson-Crick pairing or other non-traditional types.

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090130751A1 (en) * 2007-04-26 2009-05-21 Davidson Beverly L Reduction of off-target rna interference toxicity
US20100291673A1 (en) * 2006-11-29 2010-11-18 University Of Iowa Research Foundation Alternative export pathways for vector expressed rna interference
US20110111491A1 (en) * 2002-08-05 2011-05-12 University Of Iowa Research Foundation Rna interference suppresion of neurodegenerative diseases and methods of use thereof
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US8329890B2 (en) 2002-08-05 2012-12-11 University Of Iowa Research Foundation SiRNA-mediated gene silencing
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US9523093B2 (en) 2014-05-20 2016-12-20 University Of Iowa Research Foundation Huntington's disease therapeutic compounds
US11993774B2 (en) 2021-03-29 2024-05-28 Alnylam Pharmaceuticals, Inc. Huntingtin (HTT) iRNA agent compositions and methods of use thereof

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US8680063B2 (en) 2003-09-12 2014-03-25 University Of Massachusetts RNA interference for the treatment of gain-of-function disorders
WO2006121960A2 (en) 2005-05-06 2006-11-16 Medtronic, Inc. Methods and sequences to suppress primate huntington gene expression
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EP3684932A1 (en) 2017-09-22 2020-07-29 Genzyme Corporation Variant rnai

Citations (122)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4800159A (en) 1986-02-07 1989-01-24 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences
US4888829A (en) 1985-10-21 1989-12-26 Porvair Limited Gloves
US4965188A (en) 1986-08-22 1990-10-23 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme
US5236908A (en) 1991-06-07 1993-08-17 Gensia Pharmaceuticals, Inc. Methods of treating injury to the central nervous system
US5354326A (en) 1993-01-27 1994-10-11 Medtronic, Inc. Screening cable connector for interface to implanted lead
US5534350A (en) 1994-12-28 1996-07-09 Liou; Derlin Powerfree glove and its making method
US5624803A (en) 1993-10-14 1997-04-29 The Regents Of The University Of California In vivo oligonucleotide generator, and methods of testing the binding affinity of triplex forming oligonucleotides derived therefrom
US5639275A (en) 1993-08-12 1997-06-17 Cytotherapeutics, Inc. Delivery of biologically active molecules using cells contained in biocompatible immunoisolatory capsules
US5702716A (en) 1988-10-03 1997-12-30 Atrix Laboratories, Inc. Polymeric compositions useful as controlled release implants
US5720720A (en) 1993-08-27 1998-02-24 The United States Of America As Represented By The Department Of Health And Human Services Convection-enhanced drug delivery
US5735814A (en) 1996-04-30 1998-04-07 Medtronic, Inc. Techniques of treating neurodegenerative disorders by brain infusion
US5782892A (en) 1997-04-25 1998-07-21 Medtronic, Inc. Medical lead adaptor for external medical device
US5800390A (en) 1991-05-24 1998-09-01 Sumitomo Pharmaceuticals Company, Limited Equipment for intracerebral administration of preparations
US5840059A (en) 1995-06-07 1998-11-24 Cardiogenesis Corporation Therapeutic and diagnostic agent delivery
US5849995A (en) 1993-09-27 1998-12-15 The University Of British Columbia Mouse model for Huntington's Disease and related DNA sequences
US5882561A (en) 1996-11-22 1999-03-16 Drexel University Process for making a dense ceramic workpiece
US5925310A (en) 1996-03-29 1999-07-20 Asahi Glass Company Ltd. Method of making a silicon carbide product
US5942455A (en) 1995-11-14 1999-08-24 Drexel University Synthesis of 312 phases and composites thereof
US5968059A (en) 1997-03-06 1999-10-19 Scimed Life Systems, Inc. Transmyocardial revascularization catheter and method
US6042579A (en) * 1997-04-30 2000-03-28 Medtronic, Inc. Techniques for treating neurodegenerative disorders by infusion of nerve growth factors into the brain
US6093180A (en) 1995-04-28 2000-07-25 Medtronic, Inc. Intraparenchymal infusion catheter system
US6110459A (en) 1997-05-28 2000-08-29 Mickle; Donald A. G. Transplants for myocardial scars and methods and cellular preparations
US6151525A (en) 1997-11-07 2000-11-21 Medtronic, Inc. Method and system for myocardial identifier repair
US6180613B1 (en) 1994-04-13 2001-01-30 The Rockefeller University AAV-mediated delivery of DNA to cells of the nervous system
US6187906B1 (en) 1997-08-11 2001-02-13 Aukland Uniservices Limited Methods to improve neural outcome
DE19938960A1 (de) 1998-03-10 2001-02-22 Bisping Hans Juergen Testkabelanordnung
US6231969B1 (en) 1997-08-11 2001-05-15 Drexel University Corrosion, oxidation and/or wear-resistant coatings
US6245884B1 (en) 1998-10-16 2001-06-12 Vivian Y. H. Hook Secretases related to alzheimer's dementia
US6281009B1 (en) 1996-09-11 2001-08-28 The General Hospital Corporation Use of a non-mammalian DNA virus to express an exogenous gene in a mammalian cell
US6291243B1 (en) 1999-04-28 2001-09-18 The Board Of Trustees Of The Leland Stanford Jr. University P element derived vector and methods for its use
US6294202B1 (en) 1994-10-06 2001-09-25 Genzyme Corporation Compositions containing polyanionic polysaccharides and hydrophobic bioabsorbable polymers
US20010027309A1 (en) 1996-04-30 2001-10-04 Medtronic, Inc. Therapeutic method for treatment of alzheimer's disease
US6300539B1 (en) 1997-03-27 2001-10-09 Medical Research Council Model for chronic cerebral inflammation by intracerebral injection of double stranded RNA
US20010031947A1 (en) 1996-04-30 2001-10-18 Eric R. Waldkoetter Method and apparatus for drug infusion
US6310048B1 (en) 1999-12-09 2001-10-30 St. Louis University Antisense modulation of amyloid beta protein expression
US6309634B1 (en) 1998-05-27 2001-10-30 Avigen, Inc. Methods of treating Parkinson's disease using recombinant adeno-associated vector (rAAV)
US6313268B1 (en) 1998-10-16 2001-11-06 Vivian Y. H. Hook Secretases related to Alzheimer's dementia
US6319905B1 (en) 1998-12-29 2001-11-20 Cell Genesys, Inc. Method of controlling L-Dopa production and of treating dopamine deficiency
US20020004038A1 (en) 1996-04-30 2002-01-10 Baugh Robert F. Autologous platelet gel spray delivery system
US6343233B1 (en) 1997-04-25 2002-01-29 Medtronic, Inc. Medical lead adaptor
US6372250B1 (en) 2000-04-25 2002-04-16 The Regents Of The University Of California Non-invasive gene targeting to the brain
US6372721B1 (en) 1993-12-17 2002-04-16 Spinal Cord Society Method for inducing DNA synthesis in neurons
US6376471B1 (en) 1997-10-10 2002-04-23 Johns Hopkins University Gene delivery compositions and methods
US20020068093A1 (en) 2000-08-30 2002-06-06 Biocoat Incorporated Bi-laminar, hyaluronan coatings with silver- based anti-microbial properties
US6436392B1 (en) 1998-05-20 2002-08-20 University Of Iowa Research Foundation Adeno-associated virus vectors
US6436708B1 (en) 1997-04-17 2002-08-20 Paola Leone Delivery system for gene therapy to the brain
US20020114780A1 (en) 2000-11-30 2002-08-22 Krys Bankiewicz Methods of increasing distribution of therapeutic agents
US6461989B1 (en) 1999-12-22 2002-10-08 Drexel University Process for forming 312 phase materials and process for sintering the same
US6468524B1 (en) 2000-03-22 2002-10-22 The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services AAV4 vector and uses thereof
US20020187127A1 (en) 2001-04-25 2002-12-12 Krys Bankiewicz Methods of increasing distribution of nucleic acids
US6551290B1 (en) 2000-03-31 2003-04-22 Medtronic, Inc. Catheter for target specific drug delivery
US20030078229A1 (en) 2000-05-31 2003-04-24 Copernicus Therapeutics, Inc. Lyophilizable and enhanced compacted nucleic acids
US20030088236A1 (en) 1999-03-18 2003-05-08 Johnson Randolph Mellus Implantable devices and methods for treatment of pain by delivery of fentanyl and fentanyl congeners
US20030092003A1 (en) 1999-12-29 2003-05-15 Ribozyme Pharmaceuticals, Inc. Method and reagent for the treatment of Alzheimer's disease
US20030095958A1 (en) 2001-04-27 2003-05-22 Bhisetti Govinda R. Inhibitors of bace
US20030109476A1 (en) 2001-08-07 2003-06-12 Kmiec Eric B. Compositions and methods for the prevention and treatment of Huntington's disease
US20030120282A1 (en) 2001-12-24 2003-06-26 Scouten Charles W. Stereotaxic manipulator with retrofitted linear scales and digital display device
US6594880B2 (en) 1995-04-28 2003-07-22 Medtronic, Inc. Intraparenchymal infusion catheter system
US20030143732A1 (en) 2001-04-05 2003-07-31 Kathy Fosnaugh RNA interference mediated inhibition of adenosine A1 receptor (ADORA1) gene expression using short interfering RNA
US20030152947A1 (en) 2001-06-15 2003-08-14 Crossman David C. Methods for detecting and treating the early onset of aging-related conditions
US6609020B2 (en) 1999-12-01 2003-08-19 Steven Gill Neurosurgical guide device
US20030175772A1 (en) 2001-12-27 2003-09-18 Jiwu Wang Compositions for DNA mediated gene silencing
US20030190635A1 (en) 2002-02-20 2003-10-09 Mcswiggen James A. RNA interference mediated treatment of Alzheimer's disease using short interfering RNA
US6632671B2 (en) 2000-02-28 2003-10-14 Genesegues, Inc. Nanoparticle encapsulation system and method
US20030224512A1 (en) 2002-05-31 2003-12-04 Isis Pharmaceuticals Inc. Antisense modulation of beta-site APP-cleaving enzyme expression
US6659995B1 (en) 2000-11-17 2003-12-09 Syde A. Taheri Autologous myocyte micro granual retrieval and implantation (AMMGRI)
US20040018520A1 (en) 2002-04-22 2004-01-29 James Thompson Trans-splicing enzymatic nucleic acid mediated biopharmaceutical and protein
US20040023390A1 (en) 2002-08-05 2004-02-05 Davidson Beverly L. SiRNA-mediated gene silencing with viral vectors
WO2004010787A1 (ja) 2002-07-25 2004-02-05 Nissei Kabushiki Kaisha ロールコーン製造装置
US20040023855A1 (en) 2002-04-08 2004-02-05 John Constance M. Biologic modulations with nanoparticles
WO2004013280A2 (en) 2002-08-05 2004-02-12 University Of Iowa Research Foundation ALLELE-SPECIFIC siRNA-MEDIATED GENE SILENCING
WO2004047872A2 (en) * 2002-11-26 2004-06-10 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of short interfering rna (sirna)
JP2004232811A (ja) 2003-01-31 2004-08-19 Hitachi Unisia Automotive Ltd 補機駆動用電磁クラッチ
US20040186422A1 (en) 2003-03-20 2004-09-23 Robert Rioux Devices and methods for delivering therapeutic or diagnostic agents
WO2004084955A1 (en) 2003-03-20 2004-10-07 Medtronic, Inc. Anti-microbial protection for implantable medical device
US20040215164A1 (en) 2002-02-20 2004-10-28 Abbott Chun Lim Methods of treating abnormal biological conditions using metal oxides
US20040220132A1 (en) 2002-11-26 2004-11-04 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of siRNA
WO2004098648A1 (en) 2003-05-01 2004-11-18 Genzyme Corporation Gene therapy for neurometabolic disorders
WO2004101787A1 (ja) 2003-05-14 2004-11-25 Japan Science And Technology Agency ハンチンチン遺伝子の発現抑制
WO2004101063A1 (en) 2003-05-13 2004-11-25 Medtronic Inc. Medical lead adaptor assembly
US20040241854A1 (en) 2002-08-05 2004-12-02 Davidson Beverly L. siRNA-mediated gene silencing
US20040259247A1 (en) 2000-12-01 2004-12-23 Thomas Tuschl Rna interference mediating small rna molecules
US20040258666A1 (en) 2003-05-01 2004-12-23 Passini Marco A. Gene therapy for neurometabolic disorders
US20040266707A1 (en) 2003-04-02 2004-12-30 Devin Leake Stabilized polynucleotides for use in RNA interference
US20040265849A1 (en) 2002-11-22 2004-12-30 Applera Corporation Genetic polymorphisms associated with Alzheimer's disease, methods of detection and uses thereof
US20050032733A1 (en) 2001-05-18 2005-02-10 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (SiNA)
US20050042646A1 (en) 2002-08-05 2005-02-24 Davidson Beverly L. RNA interference suppresion of neurodegenerative diseases and methods of use thereof
US20050048641A1 (en) 2002-11-26 2005-03-03 Medtronic, Inc. System and method for delivering polynucleotides to the central nervous system
US6870030B2 (en) 1997-01-28 2005-03-22 Smithkline Beecham Corporation Asp2
WO2005027980A1 (en) 2003-09-12 2005-03-31 University Of Massachusetts Rna interference for the treatment of gain-of-function disorders
US20050096284A1 (en) 2002-02-20 2005-05-05 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US20050137134A1 (en) 2003-02-24 2005-06-23 North Bristol N.H.S. Trust Method of treating Parkinson's disease in humans by convection-enhanced infusion of glial cell-line derived neurotrophic factor to the putamen
US20050153353A1 (en) 2004-01-09 2005-07-14 Bernd Meibohm Real-time polymerase chain reaction-based genotyping assay for beta2-adrenergic receptor single nucleotide polymorphism
US20050191638A1 (en) 2002-02-20 2005-09-01 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US20050202075A1 (en) 2004-03-12 2005-09-15 Pardridge William M. Delivery of genes encoding short hairpin RNA using receptor-specific nanocontainers
US6945969B1 (en) 2000-03-31 2005-09-20 Medtronic, Inc. Catheter for target specific drug delivery
US20050209179A1 (en) 2000-08-30 2005-09-22 Sirna Therapeutics, Inc. RNA interference mediated treatment of Alzheimer's disease using short interfering nucleic acid (siNA)
WO2005096781A2 (en) 2004-04-06 2005-10-20 University Of Massachusetts Methods and compositions for treating gain-of-function disorders using rna interference
WO2005105995A2 (en) 2004-04-14 2005-11-10 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED TREATMENT OF POLYGLUTAMINE (POLYQ) REPEAT EXPANSION DISEASES USING SHORT INTERFERING NUCLEIC ACID (siNA)
US20050255086A1 (en) 2002-08-05 2005-11-17 Davidson Beverly L Nucleic acid silencing of Huntington's Disease gene
WO2005116204A1 (ja) 2004-05-11 2005-12-08 Rnai Co., Ltd. Ｒｎａ干渉を生じさせるポリヌクレオチド、および、これを用いた遺伝子発現抑制方法
US20050277133A1 (en) 2001-05-18 2005-12-15 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US20050282198A1 (en) 1997-05-29 2005-12-22 Interleukin Genetics, Inc. Diagnostics and therapeutics for diseases associated with an IL-1 inflammatory haplotype
US20050282188A1 (en) 2001-05-18 2005-12-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)
US20060014165A1 (en) 2003-07-14 2006-01-19 Decode Genetics Ehf. Methods of diagnosis and treatment for asthma and other respiratory diseases based on haplotype association
US20060041242A1 (en) 2001-10-31 2006-02-23 Medtronic, Inc. System and method of treating stuttering by neuromodulation
WO2006022639A1 (en) 2004-07-21 2006-03-02 Applera Corporation Genetic polymorphisms associated with alzheimer's disease, methods of detection and uses thereof
US20060150747A1 (en) 2002-07-19 2006-07-13 Phluid, Inc. Infusion pump and method for use
US20060210538A1 (en) 2004-10-22 2006-09-21 Neurologix, Inc. Use of apotosis inhibiting compounds in degenerative neurological disorders
US20060224411A1 (en) 2005-04-01 2006-10-05 Sheng-Yen Chang Method of constructing and using a memorial
WO2006121960A2 (en) * 2005-05-06 2006-11-16 Medtronic, Inc. Methods and sequences to suppress primate huntington gene expression
US20070031844A1 (en) * 2002-11-14 2007-02-08 Anastasia Khvorova Functional and hyperfunctional siRNA
WO2007039721A1 (en) 2005-10-05 2007-04-12 Astrazeneca Ab ASSOCIATION BETWEEN HLA-DRBl* 07 ALLELE AND SUSCEPTIBILY TO INCREASED LEVELS OF ALAT FOLLOWING XIMELAGATRAN ADMINISTRATION
WO2007084451A1 (en) 2006-01-17 2007-07-26 Schering Corporation Hydantoin derivatives for the treatment of inflammatory disorders
US20070184029A1 (en) 2003-12-29 2007-08-09 Am Biosolutions Method of treating cancer using platelet releasate
WO2007139811A1 (en) 2006-05-24 2007-12-06 Medtronic, Inc. Methods and kits for linking polymorphic sequences to expanded repeat mutations
US7320965B2 (en) * 2005-10-28 2008-01-22 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of Huntingtin gene
WO2008021157A1 (en) 2006-08-11 2008-02-21 Medtronic, Inc. Retrograde transport of sirna and therapeutic uses to treat neurologic disorders
WO2008046273A1 (fr) 2006-10-12 2008-04-24 Huicai Wang Machine de lavage et d'essorage constituée de balais à laver
US20090022864A1 (en) 2005-01-27 2009-01-22 Vincent Jan Steenhof Method for preparing a beverage suitable for consumption from at least two ingredients to be dissolved and/or extracted and an amount of liquid

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8274559B2 (en) * 2001-11-09 2012-09-25 Karl Storz Imaging, Inc. Replaceable hardware component of a camera control unit for video systems

Patent Citations (135)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4683202B1 (enExample) 1985-03-28 1990-11-27 Cetus Corp
US4888829A (en) 1985-10-21 1989-12-26 Porvair Limited Gloves
US4683195B1 (enExample) 1986-01-30 1990-11-27 Cetus Corp
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US4800159A (en) 1986-02-07 1989-01-24 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences
US4965188A (en) 1986-08-22 1990-10-23 Cetus Corporation Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme
US5702716A (en) 1988-10-03 1997-12-30 Atrix Laboratories, Inc. Polymeric compositions useful as controlled release implants
US5800390A (en) 1991-05-24 1998-09-01 Sumitomo Pharmaceuticals Company, Limited Equipment for intracerebral administration of preparations
US5236908A (en) 1991-06-07 1993-08-17 Gensia Pharmaceuticals, Inc. Methods of treating injury to the central nervous system
US5354326A (en) 1993-01-27 1994-10-11 Medtronic, Inc. Screening cable connector for interface to implanted lead
US5639275A (en) 1993-08-12 1997-06-17 Cytotherapeutics, Inc. Delivery of biologically active molecules using cells contained in biocompatible immunoisolatory capsules
US5720720A (en) 1993-08-27 1998-02-24 The United States Of America As Represented By The Department Of Health And Human Services Convection-enhanced drug delivery
US5849995A (en) 1993-09-27 1998-12-15 The University Of British Columbia Mouse model for Huntington's Disease and related DNA sequences
US5624803A (en) 1993-10-14 1997-04-29 The Regents Of The University Of California In vivo oligonucleotide generator, and methods of testing the binding affinity of triplex forming oligonucleotides derived therefrom
US6372721B1 (en) 1993-12-17 2002-04-16 Spinal Cord Society Method for inducing DNA synthesis in neurons
US6180613B1 (en) 1994-04-13 2001-01-30 The Rockefeller University AAV-mediated delivery of DNA to cells of the nervous system
US6294202B1 (en) 1994-10-06 2001-09-25 Genzyme Corporation Compositions containing polyanionic polysaccharides and hydrophobic bioabsorbable polymers
US5534350A (en) 1994-12-28 1996-07-09 Liou; Derlin Powerfree glove and its making method
US6594880B2 (en) 1995-04-28 2003-07-22 Medtronic, Inc. Intraparenchymal infusion catheter system
US6093180A (en) 1995-04-28 2000-07-25 Medtronic, Inc. Intraparenchymal infusion catheter system
US5997525A (en) 1995-06-07 1999-12-07 Cardiogenesis Corporation Therapeutic and diagnostic agent delivery
US5840059A (en) 1995-06-07 1998-11-24 Cardiogenesis Corporation Therapeutic and diagnostic agent delivery
US5942455A (en) 1995-11-14 1999-08-24 Drexel University Synthesis of 312 phases and composites thereof
US5925310A (en) 1996-03-29 1999-07-20 Asahi Glass Company Ltd. Method of making a silicon carbide product
US5814014A (en) 1996-04-30 1998-09-29 Medtronic Incorporated Techniques of treating neurodegenerative disorders by brain infusion
US20020004038A1 (en) 1996-04-30 2002-01-10 Baugh Robert F. Autologous platelet gel spray delivery system
US5735814A (en) 1996-04-30 1998-04-07 Medtronic, Inc. Techniques of treating neurodegenerative disorders by brain infusion
US20010031947A1 (en) 1996-04-30 2001-10-18 Eric R. Waldkoetter Method and apparatus for drug infusion
US20010027309A1 (en) 1996-04-30 2001-10-04 Medtronic, Inc. Therapeutic method for treatment of alzheimer's disease
US6281009B1 (en) 1996-09-11 2001-08-28 The General Hospital Corporation Use of a non-mammalian DNA virus to express an exogenous gene in a mammalian cell
US5882561A (en) 1996-11-22 1999-03-16 Drexel University Process for making a dense ceramic workpiece
US6870030B2 (en) 1997-01-28 2005-03-22 Smithkline Beecham Corporation Asp2
US5968059A (en) 1997-03-06 1999-10-19 Scimed Life Systems, Inc. Transmyocardial revascularization catheter and method
US6300539B1 (en) 1997-03-27 2001-10-09 Medical Research Council Model for chronic cerebral inflammation by intracerebral injection of double stranded RNA
US6436708B1 (en) 1997-04-17 2002-08-20 Paola Leone Delivery system for gene therapy to the brain
US5782892A (en) 1997-04-25 1998-07-21 Medtronic, Inc. Medical lead adaptor for external medical device
US6343233B1 (en) 1997-04-25 2002-01-29 Medtronic, Inc. Medical lead adaptor
US6042579A (en) * 1997-04-30 2000-03-28 Medtronic, Inc. Techniques for treating neurodegenerative disorders by infusion of nerve growth factors into the brain
US6110459A (en) 1997-05-28 2000-08-29 Mickle; Donald A. G. Transplants for myocardial scars and methods and cellular preparations
US20050282198A1 (en) 1997-05-29 2005-12-22 Interleukin Genetics, Inc. Diagnostics and therapeutics for diseases associated with an IL-1 inflammatory haplotype
US6187906B1 (en) 1997-08-11 2001-02-13 Aukland Uniservices Limited Methods to improve neural outcome
US6231969B1 (en) 1997-08-11 2001-05-15 Drexel University Corrosion, oxidation and/or wear-resistant coatings
US6376471B1 (en) 1997-10-10 2002-04-23 Johns Hopkins University Gene delivery compositions and methods
US6151525A (en) 1997-11-07 2000-11-21 Medtronic, Inc. Method and system for myocardial identifier repair
DE19938960A1 (de) 1998-03-10 2001-02-22 Bisping Hans Juergen Testkabelanordnung
US6436392B1 (en) 1998-05-20 2002-08-20 University Of Iowa Research Foundation Adeno-associated virus vectors
US6309634B1 (en) 1998-05-27 2001-10-30 Avigen, Inc. Methods of treating Parkinson's disease using recombinant adeno-associated vector (rAAV)
US20020141980A1 (en) 1998-05-27 2002-10-03 The Regents Of The University Of California Convection-enhanced delivery of AAV vectors
US20050180955A1 (en) 1998-05-27 2005-08-18 Regents Of The University Of California Methods of treating parkinson's disease using viral vectors
US6313268B1 (en) 1998-10-16 2001-11-06 Vivian Y. H. Hook Secretases related to Alzheimer's dementia
US6245884B1 (en) 1998-10-16 2001-06-12 Vivian Y. H. Hook Secretases related to alzheimer's dementia
US6319905B1 (en) 1998-12-29 2001-11-20 Cell Genesys, Inc. Method of controlling L-Dopa production and of treating dopamine deficiency
US20030088236A1 (en) 1999-03-18 2003-05-08 Johnson Randolph Mellus Implantable devices and methods for treatment of pain by delivery of fentanyl and fentanyl congeners
US6291243B1 (en) 1999-04-28 2001-09-18 The Board Of Trustees Of The Leland Stanford Jr. University P element derived vector and methods for its use
US6609020B2 (en) 1999-12-01 2003-08-19 Steven Gill Neurosurgical guide device
US6310048B1 (en) 1999-12-09 2001-10-30 St. Louis University Antisense modulation of amyloid beta protein expression
US6461989B1 (en) 1999-12-22 2002-10-08 Drexel University Process for forming 312 phase materials and process for sintering the same
US20030092003A1 (en) 1999-12-29 2003-05-15 Ribozyme Pharmaceuticals, Inc. Method and reagent for the treatment of Alzheimer's disease
US6632671B2 (en) 2000-02-28 2003-10-14 Genesegues, Inc. Nanoparticle encapsulation system and method
US6468524B1 (en) 2000-03-22 2002-10-22 The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services AAV4 vector and uses thereof
US6945969B1 (en) 2000-03-31 2005-09-20 Medtronic, Inc. Catheter for target specific drug delivery
US6551290B1 (en) 2000-03-31 2003-04-22 Medtronic, Inc. Catheter for target specific drug delivery
US6372250B1 (en) 2000-04-25 2002-04-16 The Regents Of The University Of California Non-invasive gene targeting to the brain
US20030078229A1 (en) 2000-05-31 2003-04-24 Copernicus Therapeutics, Inc. Lyophilizable and enhanced compacted nucleic acids
US20050209179A1 (en) 2000-08-30 2005-09-22 Sirna Therapeutics, Inc. RNA interference mediated treatment of Alzheimer's disease using short interfering nucleic acid (siNA)
US20020068093A1 (en) 2000-08-30 2002-06-06 Biocoat Incorporated Bi-laminar, hyaluronan coatings with silver- based anti-microbial properties
US6659995B1 (en) 2000-11-17 2003-12-09 Syde A. Taheri Autologous myocyte micro granual retrieval and implantation (AMMGRI)
US20020114780A1 (en) 2000-11-30 2002-08-22 Krys Bankiewicz Methods of increasing distribution of therapeutic agents
US20040259247A1 (en) 2000-12-01 2004-12-23 Thomas Tuschl Rna interference mediating small rna molecules
US20030143732A1 (en) 2001-04-05 2003-07-31 Kathy Fosnaugh RNA interference mediated inhibition of adenosine A1 receptor (ADORA1) gene expression using short interfering RNA
US20020187127A1 (en) 2001-04-25 2002-12-12 Krys Bankiewicz Methods of increasing distribution of nucleic acids
US20030095958A1 (en) 2001-04-27 2003-05-22 Bhisetti Govinda R. Inhibitors of bace
US20050032733A1 (en) 2001-05-18 2005-02-10 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (SiNA)
US20050277133A1 (en) 2001-05-18 2005-12-15 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US20050282188A1 (en) 2001-05-18 2005-12-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)
US20030152947A1 (en) 2001-06-15 2003-08-14 Crossman David C. Methods for detecting and treating the early onset of aging-related conditions
US20030109476A1 (en) 2001-08-07 2003-06-12 Kmiec Eric B. Compositions and methods for the prevention and treatment of Huntington's disease
US20060041242A1 (en) 2001-10-31 2006-02-23 Medtronic, Inc. System and method of treating stuttering by neuromodulation
US20030120282A1 (en) 2001-12-24 2003-06-26 Scouten Charles W. Stereotaxic manipulator with retrofitted linear scales and digital display device
US20030175772A1 (en) 2001-12-27 2003-09-18 Jiwu Wang Compositions for DNA mediated gene silencing
US20050191638A1 (en) 2002-02-20 2005-09-01 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US20040215164A1 (en) 2002-02-20 2004-10-28 Abbott Chun Lim Methods of treating abnormal biological conditions using metal oxides
US20050096284A1 (en) 2002-02-20 2005-05-05 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US20030190635A1 (en) 2002-02-20 2003-10-09 Mcswiggen James A. RNA interference mediated treatment of Alzheimer's disease using short interfering RNA
US20040023855A1 (en) 2002-04-08 2004-02-05 John Constance M. Biologic modulations with nanoparticles
US20040018520A1 (en) 2002-04-22 2004-01-29 James Thompson Trans-splicing enzymatic nucleic acid mediated biopharmaceutical and protein
US20030224512A1 (en) 2002-05-31 2003-12-04 Isis Pharmaceuticals Inc. Antisense modulation of beta-site APP-cleaving enzyme expression
US20060150747A1 (en) 2002-07-19 2006-07-13 Phluid, Inc. Infusion pump and method for use
WO2004010787A1 (ja) 2002-07-25 2004-02-05 Nissei Kabushiki Kaisha ロールコーン製造装置
US20050106731A1 (en) 2002-08-05 2005-05-19 Davidson Beverly L. siRNA-mediated gene silencing with viral vectors
US20040023390A1 (en) 2002-08-05 2004-02-05 Davidson Beverly L. SiRNA-mediated gene silencing with viral vectors
US20050255086A1 (en) 2002-08-05 2005-11-17 Davidson Beverly L Nucleic acid silencing of Huntington's Disease gene
US20050042646A1 (en) 2002-08-05 2005-02-24 Davidson Beverly L. RNA interference suppresion of neurodegenerative diseases and methods of use thereof
US20040241854A1 (en) 2002-08-05 2004-12-02 Davidson Beverly L. siRNA-mediated gene silencing
US20060009408A1 (en) * 2002-08-05 2006-01-12 University Of Iowa Research Foundation, A Iowa Corporation siRNA-Mediated gene silencing with viral vectors
WO2004013280A2 (en) 2002-08-05 2004-02-12 University Of Iowa Research Foundation ALLELE-SPECIFIC siRNA-MEDIATED GENE SILENCING
WO2004013355A1 (en) 2002-08-05 2004-02-12 University Of Iowa Research Foundation Sirna-mediated gene silencing with viral vectors
US20070031844A1 (en) * 2002-11-14 2007-02-08 Anastasia Khvorova Functional and hyperfunctional siRNA
US20040265849A1 (en) 2002-11-22 2004-12-30 Applera Corporation Genetic polymorphisms associated with Alzheimer's disease, methods of detection and uses thereof
US20050048641A1 (en) 2002-11-26 2005-03-03 Medtronic, Inc. System and method for delivering polynucleotides to the central nervous system
WO2004047872A2 (en) * 2002-11-26 2004-06-10 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of short interfering rna (sirna)
US20040220132A1 (en) 2002-11-26 2004-11-04 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of siRNA
JP2004232811A (ja) 2003-01-31 2004-08-19 Hitachi Unisia Automotive Ltd 補機駆動用電磁クラッチ
US20050137134A1 (en) 2003-02-24 2005-06-23 North Bristol N.H.S. Trust Method of treating Parkinson's disease in humans by convection-enhanced infusion of glial cell-line derived neurotrophic factor to the putamen
US20040186422A1 (en) 2003-03-20 2004-09-23 Robert Rioux Devices and methods for delivering therapeutic or diagnostic agents
WO2004084955A1 (en) 2003-03-20 2004-10-07 Medtronic, Inc. Anti-microbial protection for implantable medical device
US20040266707A1 (en) 2003-04-02 2004-12-30 Devin Leake Stabilized polynucleotides for use in RNA interference
WO2004098648A1 (en) 2003-05-01 2004-11-18 Genzyme Corporation Gene therapy for neurometabolic disorders
US20040258666A1 (en) 2003-05-01 2004-12-23 Passini Marco A. Gene therapy for neurometabolic disorders
WO2004101063A1 (en) 2003-05-13 2004-11-25 Medtronic Inc. Medical lead adaptor assembly
WO2004101787A1 (ja) 2003-05-14 2004-11-25 Japan Science And Technology Agency ハンチンチン遺伝子の発現抑制
US20060014165A1 (en) 2003-07-14 2006-01-19 Decode Genetics Ehf. Methods of diagnosis and treatment for asthma and other respiratory diseases based on haplotype association
WO2005027980A1 (en) 2003-09-12 2005-03-31 University Of Massachusetts Rna interference for the treatment of gain-of-function disorders
US20070184029A1 (en) 2003-12-29 2007-08-09 Am Biosolutions Method of treating cancer using platelet releasate
US20050153353A1 (en) 2004-01-09 2005-07-14 Bernd Meibohm Real-time polymerase chain reaction-based genotyping assay for beta2-adrenergic receptor single nucleotide polymorphism
US20050202075A1 (en) 2004-03-12 2005-09-15 Pardridge William M. Delivery of genes encoding short hairpin RNA using receptor-specific nanocontainers
US20050288243A1 (en) 2004-04-06 2005-12-29 University Of Massachusetts Methods and compositions for treating gain-of-function disorders using RNA interference
WO2005096781A2 (en) 2004-04-06 2005-10-20 University Of Massachusetts Methods and compositions for treating gain-of-function disorders using rna interference
GB2415961A (en) 2004-04-14 2006-01-11 Sirna Therapeutics Inc RNA interference (RNAi) of huntingtin (htt) gene
WO2005105995A2 (en) 2004-04-14 2005-11-10 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED TREATMENT OF POLYGLUTAMINE (POLYQ) REPEAT EXPANSION DISEASES USING SHORT INTERFERING NUCLEIC ACID (siNA)
WO2005116204A1 (ja) 2004-05-11 2005-12-08 Rnai Co., Ltd. Ｒｎａ干渉を生じさせるポリヌクレオチド、および、これを用いた遺伝子発現抑制方法
US20080113351A1 (en) * 2004-05-11 2008-05-15 Alphagen Co., Ltd. Polynucleotides for causing RNA interference and method for inhibiting gene expression using the same
WO2006022639A1 (en) 2004-07-21 2006-03-02 Applera Corporation Genetic polymorphisms associated with alzheimer's disease, methods of detection and uses thereof
US20060210538A1 (en) 2004-10-22 2006-09-21 Neurologix, Inc. Use of apotosis inhibiting compounds in degenerative neurological disorders
US20090022864A1 (en) 2005-01-27 2009-01-22 Vincent Jan Steenhof Method for preparing a beverage suitable for consumption from at least two ingredients to be dissolved and/or extracted and an amount of liquid
US20060224411A1 (en) 2005-04-01 2006-10-05 Sheng-Yen Chang Method of constructing and using a memorial
US20060257912A1 (en) 2005-05-06 2006-11-16 Medtronic, Inc. Methods and sequences to suppress primate huntington gene expression
WO2006121960A2 (en) * 2005-05-06 2006-11-16 Medtronic, Inc. Methods and sequences to suppress primate huntington gene expression
WO2007039721A1 (en) 2005-10-05 2007-04-12 Astrazeneca Ab ASSOCIATION BETWEEN HLA-DRBl* 07 ALLELE AND SUSCEPTIBILY TO INCREASED LEVELS OF ALAT FOLLOWING XIMELAGATRAN ADMINISTRATION
US7320965B2 (en) * 2005-10-28 2008-01-22 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of Huntingtin gene
WO2007084451A1 (en) 2006-01-17 2007-07-26 Schering Corporation Hydantoin derivatives for the treatment of inflammatory disorders
WO2007139811A1 (en) 2006-05-24 2007-12-06 Medtronic, Inc. Methods and kits for linking polymorphic sequences to expanded repeat mutations
WO2008021157A1 (en) 2006-08-11 2008-02-21 Medtronic, Inc. Retrograde transport of sirna and therapeutic uses to treat neurologic disorders
WO2008046273A1 (fr) 2006-10-12 2008-04-24 Huicai Wang Machine de lavage et d'essorage constituée de balais à laver

Non-Patent Citations (205)

* Cited by examiner, † Cited by third party
Title
Aebisher, Trends in Neurosci. 24(9) 553-540 (Sep. 2001).
Altschul et al., "Gapped BLAST and PSO-BLAST: a new generation of protein database search prorams," Nucl. Acids Res., 25(17): 3389-3402 (1997).
Ambion Inc., pSilencer(TM) 1.0-U6 siRNA Expression Vector, Catalog # 7207-20 mug, Nov. 2004, Austin, TX, 6 pgs.
Ambion Inc., pSilencer™ 1.0-U6 siRNA Expression Vector, Catalog # 7207-20 μg, Nov. 2004, Austin, TX, 6 pgs.
Ambion Technical Bulletin #506 (as published on Nov. 16, 2002) downloaded from www.archive.org.
Ambion, Inc., Silencer siRNA® Construction Kit, Cat. #1620, Instruction Manual, Aug. 2005, 36 pgs.
Ausubel et al., Eds., Current Protocols in Molecular Biology, vols. 1-3, John Wiley & Sons, Inc., New York, NY, 1994; title page, publisher's page and table of contents only, 14 pgs.
Basi et al., "Antagonistic Effects of beta-site Amyloid Precursor Protein-cleaving Enzymes 1 and 2 on beta- Amyloid Peptide Production in Cells," J. Bio. Chem., Published, JBC Papers in Press, Jun. 2003; 278(34): 31512-31520.
Basi et al., "Antagonistic Effects of β-site Amyloid Precursor Protein-cleaving Enzymes 1 and 2 on β- Amyloid Peptide Production in Cells," J. Bio. Chem., Published, JBC Papers in Press, Jun. 2003; 278(34): 31512-31520.
Bass et al., Nature 411: 428-429 (2001).
Bertrand et al., Biochem Biophys Res Comm 296: 1000-1004 (2002).
Bodendorf et al., J. Neurochem. 80(5), 799-806 (Mar. 2002).
Boillée et al., "Gene therapy for ALS deliver," Trends in Neurosciences, May 2004; 27(5): 235-238.
Bortolin, Susan et al., "Analytical validation of the tag-it high-throughout microsphere-based universal array genotyping platform. Application to the multiplex detection of a panel of thrombophilia-associates single-nucleotide polymorphisms." American Association for Clinical Chemistry vol. 50(11) 2028-2036 (2004).
Brentano et al., P.N.A.S. 89:4099-4103 (1992).
Brummelkamp et al., Science 296: 550-553 (2002).
Burger et al., Mol. Ther. 10(2) 302-317 (Aug. 2004).
Cahill et al Atlas of Human Cross-Sectional Anatomy Wiley-Liss, 3rd Ed. (1995).
Cai et al., Nat. Neurosci. 4(3) 233-234 (2004).
Callahan Am. J. Pathol. 158(3) 1173-1177 (2001).
Caplen et al, Human Mol. Genet. 11(2) 175-184 (2002).
Chen et al., Nucl. Acid. Res. 20, 4581-4589 (1992).
Chi et al., "Genomewide view of gene silencing by small interfering RNAs," Proc. Natl. Acad. Sci. USA, May 2003; 100 (11): 6343-6346.
Chowhira et al., J. Biol. Chem. 269, 25856-25863 (1994).
Christman, Tissue Engineering (10) 403-409 (2004).
Cioffi et al., Biochem J. 365: 833-840 (2002).
Clark et al., Annals Int. Med. 138 400-411 (2003).
Clark et al., J. Neurosci. 17(19) 7385-7395 (1997).
Cleary et al., Nat. Neurosci. 8(1) 79-84 (ePub Dec. 19, 2004).
Couture et al., Trends in Genetics, 12(12) 510-515 (Dec. 1996).
Dai et al., Developmental Biology 285:80-90 (2005).
Davidson et al., The Lancet, Neurology 3, 145-149 (2004).
Demetriades J. Neurolog. Sci. 203-204, 247-251 (2002).
Dineley, J, Biol. Chem. 277 (25) 22768-22780 (2002).
Dorri et al., Exp. Neurology 147 48-54 (1997).
Dropulic et al., J. Virol. 66(1) 1432-1441 (1992).
During et al., "Subthalamic GAD GeneTransfer in Parkinson's Disease Patients Who Are Candidates for Deep Brain Stimulation," Human Gene Therapy, Aug. 2001; I2(12): 1587-1598.
Elbashir et al., "Analysis of gene function in somatic mammalian cells using small interfering RNAs," Methods 26 (2002); pp. 199-213 I 1.
ElBashir, EMBOJ 20(23) 6877-6888 (2001).
Ezrin-Waters et al., Can. J. Neurol. Sci. 13, 8-14 (1986).
Fu et al., Mo. Ther. 8(6) 911-917 (Dec. 2003).
Gau, Am. J. Pathol., 160(2) 731-738 (2002).
GeneDetect.com Limited, Code GD100X-RV, (GeneDetect rAVE(TM) gene delivery reagent), copyright 2000-2002, Auckland, New Zealand, 2 pgs.
GeneDetect.com Limited, Code GD100X-RV, (GeneDetect rAVE™ gene delivery reagent), copyright 2000-2002, Auckland, New Zealand, 2 pgs.
Geraerts et al., Concise Review: Therapeutic Strategies for Parkinson Disease Based on Modulation of Adult Neurogenesis. Stem Cells, Nov. 2, 2006, vol. 25, No. 2, pp. 263-270.
Gerlai Behav. Brain Res. 95 191-203 (1998).
Glorioso, Curr. Opinion in Drug Discovery & Dev't 5(2) Pharma Press ISSN 1367-6733 (2002).
Good et al., Gene Ther. 4: 45-54 (1997).
Goto et al., Neurology, 60(5) Suppl. 1 p. A286 (Mar. 11, 2003).
Harrison et al., Mol. Cel. Neurosci. 24(3) 646-655 (2003).
Hartlage-Rubsamen et al., Glia 41(2) 169-179 (Dec. 28, 2002).
Heale et al., Nucl. Acid. Res. 22(3), 2005.
Holen et al., Nucl. Acid Res. 30:1757-1766 (2002).
Hommel et al., "Local gene knockdown in the brain using viral-mediated RNA interference," Nature Medicine, Dec. 2003; 9(12); 1539-1544.
Hommel et al., Society for Neuroscience Abstract, 2003, Abstract 325.14 (2003).
Hooper et al., Neuroscience 63, 917-924 (1995).
Hsiao et al, Science 274 99-102(1996).
Huwylet et al., "Brain drug delivery of small molecules using immunoliposomes,"Proc. Natl. Acad., USA, Nov. 1996;93:14164-14169.
Icascon et al., Scandinavian Physiol. Society 179 173-177 (2003).
Invitrogen, pShooter(TM) Vector (pCMV/myc® vectors), For the intracellular targeting of recombinant proteins and antibodies, Catalog Nos. V820-20, V821-20, V822-20, V823-20, Version E, copyright 1998-2001, 35 pgs.
Invitrogen, pShooter™ Vector (pCMV/myc® vectors), For the intracellular targeting of recombinant proteins and antibodies, Catalog Nos. V820-20, V821-20, V822-20, V823-20, Version E, copyright 1998-2001, 35 pgs.
Invitrogen, pTRACERS(TM) -CMV2, Catalog Nos. V885-01, V885-20, Version C, copyright 1998-2001, 21 pgs.
Invitrogen, pTRACERS™ -CMV2, Catalog Nos. V885-01, V885-20, Version C, copyright 1998-2001, 21 pgs.
Izant et al., Science 299 345 (1985).
Kaemmerer et al., Soc. Neurosci. Meeting (Oct. 26, 2004).
Kao et al., "BACE1 Suppression by RNA Interference in Primary Cortical Neurons," J. Bio. Chem., Published, JBC Papers inPress, Nov. 2003, 2004; 279(3): 1942-1949.
Kashani-Sabet et al., Antisense Res. Dev. 2: 3-15 (1992).
Katz et al., Bioessays 11(6): 181-185 (Dec. 1989).
Kawarabayashi et al., J. Neurosci. 372-381 (2001).
Kenderell et al., (2000) Nat. Biotech. 17, 896-898 (2000).
King et al., Physiology & Behavior, 75:627-642, 2002.
Kitabwala et al., New England J. Med. 347(17) 1364-1367 (Oct. 24, 2002).
Kitazume J. Biol. Chem. 280(9) 8589-8595 (Mar. 4, 2005).
Klement et al., Cell 95 41-53 (1998).
Laird et al., J. Neurosci. 25, 11693-11709 (Dec. 14, 2005).
Le Gal La Salle et al, Science 259, 988-990 (1993).
L'Huillier et al., EMBO J. 11(12), 4411-4418 (1992).
Li et al., "Predicting siRNA efficiency," Cell. Mol. Life Sci. 64 (2007), pp. 1785-1792.
Li et al., Mol. Cell Bio. 22 (5) 1277-1287 (2002).
Lisziewicz et al., Proc. Nat. Acad. Sci 90 8000-8004 (Sep. 1993).
Liu et al., Proc. Japan Academy, Series B, Physical and Biol. Sciences 79(10) 293-298 (Dec. 1993).
Luo et al., Neurobiol. Dis. 14(1), 81-88 (Oct. 2003).
Luo, Nat. Neurosci. 4, 231-232 (2001).
MacDonald, M. et al., "A novel gene containing a trinucleotide repeat that is expanded and unstable on huntington's disease chromosomes," Cell, vol. 72, 971-983 (1993).
Mas-Monteys, A. et al., "Allele-Specific silencing of mutant huntingtin for huntington's disease therapy", Molecular Therapy 13: S274-S275 (2006).
Matilla et al., J. Neurosci 18, 5508-5516 (1998).
McGarry et al., Proc. Natl Acad. Sci. 83, 399-403 (1986).
McManus et al., Nature Reviews/Genetics 3, 737-747 (Oct. 2002).
Menei et al Neurosurgery 34: 1058-1064 (1994).
Messier et al., Pharm. Biochem Behavior 63 313-318 (1999).
Miller et al Proc. Nat. Acad. Sci. 100(12) 7195-7200 (Jun. 10, 2003).
Mogan et al., JECT 36: 191-196 (2004).
Morel et al., J. Comparative Neurology 387, 588-630 (1997).
Mucke et al., J. Neurosci. 20(11) 4050-4058 (Jun. 1, 2000).
Naldini et al., Proc. Nat. Acad. Sci. 93, 11382-11388 (Oct. 1996).
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, "What does NCBI do?" [online]. Bethesda, MD [retrieved on Dec. 5, 2009], Revised Dec. 2005. Retrieved from the Internet: ; 2 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, "What does NCBI do?" [online]. Bethesda, MD [retrieved on Dec. 5, 2009], Revised Dec. 2005. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.gov>; 2 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus AF163864, Accession No. AF163864, "Homo sapiens SNCA isoform (SNCA) gene, complete cds, alternatively spliced, " [online]. Bethesda, MD [retrieved on Jun. 21, 2004]. Retrieved from the Internet: ; 43 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus AF163864, Accession No. AF163864, "Homo sapiens SNCA isoform (SNCA) gene, complete cds, alternatively spliced, " [online]. Bethesda, MD [retrieved on Jun. 21, 2004]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=11118351>; 43 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus AF163865, Accession No. AF163865, "Mus musculus alpha-synuclein (Snca) gene, complete cds," [online]. Bethesda, MD [retrieved on Jun. 21, 2004]. Retrieved from the Internet; 33 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus AF163865, Accession No. AF163865, "Mus musculus alpha-synuclein (Snca) gene, complete cds," [online]. Bethesda, MD [retrieved on Jun. 21, 2004]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=11118354>; 33 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus AH003045, Accession No. AH003045, "Homo sapiens huntingtin (HD) gene, exon 1," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the internet:; 42 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus AH003045, Accession No. AH003045, "Homo sapiens huntingtin (HD) gene, exon 1," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the internet:<URL:www.ncbi.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=663286>; 42 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000027, Acession No. NM—000027, "Homo sapiens aspartylgluocsaminidase (AGA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=38569404>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM-000027, Acession No. NM-000027, "Homo sapiens aspartylgluocsaminidase (AGA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM-000046, Accession No. NM-000046, "Homo sapiens arylsulfatase B (ARSB), transcript variant 1, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieeved from the Internet:URL; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000046, Accession No. NM—000046, "Homo sapiens arylsulfatase B (ARSB), transcript variant 1, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieeved from the Internet:URL<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4557334>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM-000049, Accession No. NM-000049, "Homo sapiens aspartoacylase (aminoacylase 1, Canavan disease). (ASPA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:; 3 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000049, Accession No. NM—000049, "Homo sapiens aspartoacylase (aminoacylase 1, Canavan disease). (ASPA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=24475878>; 3 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—0001147, Accession No. NM—000147, "Homo sapiens fucosidase, alpha-L1, acid tissue (FUCA1), mRNA" [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=24475878>; 3 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000152, Accession NM—000152, "Homo sapiens glucosidase, alpha; acid (Pompe disease, glycogen storage disease type II) (GAA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=11496988>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000153, Accession No. NM—000153, "Homo sapiens galactosylceramidase (Krabbe disease) (GALC), mRNA," [online]. Bethesda, MD [retrieved Mar. 5, 2009]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4557612>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000157, Accession No. NM—000157, "Homo sapiens glucodase, beta; acid (includes glucosylceramidase) (GBA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4503934>; 7 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000157, Accession No. NM—000158, "Homo sapiens glucan (1, 4-alpha-), branching enzyme 1 (glucogen branching enzyme, Andersen disease, glycogen storage disease type (IV)(GBE1)," [online]. Bethesda, MD db=nucleotide&val=4557618>; 2 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000181, Accession No. NM—000181, "Homo sapiens glucoronidase, beta (GUSB), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=450422>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000199, Accession No. NM—000199, "Homo sapiens N-sulfoglucosamine sulfohydrolase (sulfamidase) (SGSH), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=31543619>; 3 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000202, Accession No. NM—000202, "Homo sapiens iduronate 2-sulfatase (Hunter syndrome)(ID), transcript variant 1, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=5360215>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000235, Accession No. NM—000235, "Homo sapiens lipase A, lysosomal acid, cholesterol esterase (Wolman disease) (LIPA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4557720>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000262, Accession No. NM—000262, "Homo sapiens N-acetylgalactosaminidase, alpha- (NAGA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4557780>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000263, Accession No. NM—000263, "Homo sapiens N-acetylglucosaminidase, alpha-(Sanfilippo disease) (IIIB)(NAGLU), mRNA," [online]. Bethesda, MD [retrieved on Mar. 3, 2009]. Retrieved from the Internet<URL:http:// www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=40548380>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000310, Accession No. NM—000310, "Homo sapiens palmitoyl-protein thioesterase 1 (ceroid-lipofuscinosis, neuronal 1, infantile) (PPT1), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved froM the Internet: <URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4506030>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000332, Accession No. NM—000332, "Homo sapiens spinocerebellar ataxia 1 (olivopontocerebellar ataxia 1, autosomal dominant, ataxin 1) (SCA1), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4506792>; 7 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000345, Accession No. NM—000345, "Homo sapiens synuclein, alpha (nonA4 component of amyloid precursor) (SNCA), transcript variant NACP140, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=6806896>; 6 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000404, Accession No. NM—000404, "Homo sapiens glactosidase, beta 1 (GLB1), tranascript variant 179423, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Interna<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=10834965>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000434, Accession No. NM—000434, "Homo sapiens sialidase 1 (lysosomal sialidase)(NEU1), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=40806202>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000487, Accession No. NM—000487, "Homo sapiens arysulfatase A (ARSA), mRNA," 1 MD [retrieved on 2009-03-05]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi? db=nucleotide&val=7262293>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000512, Accession No. NM—000512, "Homo sapiens galactosamine (N-acetyl)-6-sulfate sulfatase (Morquio syndrome, mucopolysaccharidosis type IVA), (GALNS), mRNA," [online]. Bethesda, MD [retrieved Mar. 5, 2009]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih/gov/entrez/viewer.fcgi?db=nucleotides&val=9945384>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000520, Accession No. NM—000520, "Homo sapiens hexosamidase A (alpha polypeptide) (HEXA), mRNA,"[online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=13128865>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000521, Accession No. NM—000521, "Homo sapiens hexosaminidase B (beta polypeptide) (HEXB), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=13128866>; 6 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000528, Accession No. NM—000528, "Homo sapiens mannosidase, alpha, class 2B, member 1 (MAN2B1), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=10834967>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—000543, Accession No. NM—000543, "Homo sapiens sphingomyelin phosphodiesterase 1 acid lysosomal (acid sphingomyelinase) (SMPD1), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=40254417>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—002076, Accession No. NM—002076, "Homo sapiens glucosamine (N-acetyl)-6-sulfatase (Sanfilippo disease)(IIId)(GNS), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL: http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=42490755>; 7 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—002778, Accession No. NM—0002778, Accession No. NM—000169, "Homo sapiens glactosidase, alpha (GLA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=4504008>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—002778, Accession No. NM—002778, "Homo sapiens prosaposin (variant Gaucher disease and variant metachromatic leukodystrophy) (PSAP), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=11386146>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—004315, Accession No. NM—004315, "Homo sapiens N-acylsphingosine amidohydrolase (acid ceramidase) 1 (ASAHI), transcript variant 2, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=30089929>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—004993, Accession No. NM—004993,"Homo sapiens Machado-Joseph disease (spinocerebellar ataxia 3, olivopontocerebellar ataxia 3, autosomal dominant, ataxin 3) (MJD), transcript variant 1, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi? db=nucleotide&val=13518018>; 9 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—005908, Accession No. NM—005908, "Homo sapiens mannosidase, beta A, lyosomal (MANBDA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=24797157>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—007308, Accession No. NM—007308, "Homo sapiens synuclein, alpha (nonA4 component of amyloid precursor) (SNCA), transcript variant NACP112, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=6806897>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—009124, Accession No. NM—009124, "Definition," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=33636695>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—011792, Accession No. NM—011792, Version NM—011792.2, "Mus musculus beta-site APP cleaving enzyme 1 (BACE 1), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http:// www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=31981411>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—011792.2, Accession No. NM—011792, "Mus musculus beta-site APP cleaving enzyme (BACE), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbionlmonih.gov/entrez/viewer.fcgi?db=nucleotide&val=6857758>; 4 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—012104, Accession No. NM—012104, "Homo sapiens beta-site APP-cleaving enzyme 1 (BACE1), transcript variant a, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=46255011>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—012104, Accession No. NM—012104, Version NM—012104.2, "Homo sapiens beta-site APP-cleaving enzyme (BACE), transcript variant a, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=21040369>; 10 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—013995, Accession No. NM—013995, "Homo sapiens lysosomal-associated membrane protein 2 (LAMP2), transcript variant LAMP2B, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet: <URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=7669502>; 6 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—030660, Accession No. NM—030660, "Homo sapiens Machado-Joseph disease (spinocerebellar ataxia 3, olivopontocerebellar ataxia 3, autosomal dominant, ataxin 3) (MJD), transcript variant 2, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi? db=nucleotide&val=13518012>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—032520, Accession No. NM—032520, "Homo sapiens N-acetylglucosamine-1-phosphotransferase, gamma subunit (GNPTAG), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.govientrez/viewer.fcgi?db=nucleotide&val=42476109>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—138971, Accession No. NM—138971, "Homo sapiens beta-site APP-cleaving enzyme 1 (BACE1), transcript variant c, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=46255012>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—138971, Accession No. NM—138971, Version NM—138971.1, "Homo sapiens beta-site APP-cleaving enzyme (BACE), transcript variant c, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=21040363>; 10 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—138972, Accession No. NM—138972, "Homo sapiens beta-site APP-cleaving enzyme 1 (BACE1), transcript variant b, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=46255013>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—138972, Accession No. NM—138972, Version NM—138972.1, "Homo sapiens beta-site APP-cleaving enzyme (BACE), transcript variant b, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=21040365>; 10 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—138973, Accession No. NM—138973, "Homo sapiens beta-site APP-cleaving enzyme 1 (BACE1), transcript variant d, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet:<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=46255014>; 8 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus NM—138973, Accession No. NM—138973, Version NM—138973.1, "Homo sapiens beta-site APP-cleaving enzyme (BACE), transcript variant d, mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=21040367>; 10 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus U24233, Accession No. U24233, "Mus musculus huntingtin (Hd) mRNA, complete cds," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi? db=nucleotide&val=902003>; 5 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus XM—032588, Accession No. XM—032588, "Homo sapiens dentatorubral-pallidoluysian atrophy (artrophin-1) (DRPLA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=20555988>; 3 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank Locus XM—132846, Accession No. XM—132846, "Mus musculus dentatorubral pallidoluysian atrophy (DRPLA) mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbionlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=20832263>; 3 pgs.
National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, GenBank.Locus NM—000203, Accession No. NM—000203, "Homo sapiens iduronidase, alpha-L-(IDUA), mRNA," [online]. Bethesda, MD [retrieved on Mar. 5, 2009]. Retrieved from the Internet<URL:http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=40354208>; 6 pgs.
Noonberg et al., Nucl. Acid Res. 22(14) 2830-2836 (1994).
Noordmans et al., Soc. Neurosci. Abstr. 27, Program 572.14 (2001).
Ohkawa Nucl. Acid. Symp. Ser. 27, 15-16 (1992).
Ohno et al., "BACEI Deficiency Rescues Memory Deficits and Cholinergic Dysfunction in a Mouse Model of Alzheimer's Disease," Neuron, Jan. 2004; 41: 27-33.
Paxinos et al The Mouse Brain in Stereotactic Coordinates, Acad. Press 2nd Ed. (2001).
Potter, N. T. et al., "Technical standards and guidelines for huntington disease testing," Genetics in Medicine 6:61-65 (2004).
Qiagen, Rneasy Mini Handbook, 3rd Edition, Jun. 2001, 116 pgs.
Qiangen, Qiaex II Handbook, Feb. 1999, 24 pgs.
Roberds et al., "BACE knockout mice are healthy despite lacking the primary β-secretase activity in brain: implications for Alzheimer's disease therapeutics," Human Molecular Genetics, Jun. 2001; 10(12): 1317-1324.
Ryu, Biomaterials 26: 319-326 (2005).
Salehi et al., J. Neural Transm. 106 955-986 (1999).
Sapru et al., Annual Meeting Soc. Neurosci. Abstract 297.9, XP001204566 (2003).
Sarver et al., Science 247, 1222-1225 (1990).
Scanlon et al., Proc. Nat. Acad. Sci. 88, 10591-10595 (1995).
Schenk, "Amyloid-β immunotherapy for Alzheimer's disease: the end of the beginning," Nature Reviews—Neuroscience, Oct. 2002; 3: 824-828.
Scherr et al., Cell Cycle 2(3) 251-257 (2003).
Serra et at., Medical Image Analysis 1(4) 317-329 (1996).
Singer et al., Nat. Neurosci. 8(10) 1343-1349 (ePub Aug. 28, 2005) .
Stackman et al., Experimental Neurology 184, 510-520 (2003).
Strategene, AAV Helper-Free System, Instruction Manual, Catalog #240071, #240074, #240075, Revision #084007i, Aug. 2004, 50 pgs.
Strategene, pBluescript® II Phagemid Vectors, Instruction Manual, Catalog #212205, #212206, #212207, #212208, Revision #083001m, Aug. 2003, 35 pgs.
Sullenger, Science 262, p. 1566 (12.03.1993).
Taira et al., Nucl. Acid. Res. 19(19) 5125-5130 (1991).
Thompson et al., Nucl. Acid. Res. 23(12), 2259 (1995).
Timson et al., Biochem J 363:515-520 (2002).
Tuscjl Lab, "The siRNA user guide," Revised May 2004, [online]. Retrieved on Nov. 29, 2005. Retrieved from the Internet: <URL:rockefelleredu/labheads/tuschl/sirna.html>, 6 pgs.
Ui-Tei et al (Nucleic Acids Research, 2004; vol. 34, No. 3, pp. 936-948). *
Ui-Tei et al., "Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference,"; Nucleic Acids Research (2004); vol. 32, No. 3, pp. 936-948.
Valbonesi et al., Ttransf. And Apheresis Sci. 30: 153-156 (2004).
Van Bilsen et al., "Identification and allele-specific silencing of the mutant huntingtin allele in Huntington's disease patient-derived fibroblasts," Human Gene Therapy, vol. 19, pp. 710-718 (2008).
Vassar et al., Science 286 735-741 (1999).
Ventura et al., Nucl. Acid. Res. 21(14) 3249-3255 (1993).
Vickers, Journal of Bio. Chemistry, vol. 278, No. 9 7108-7118 (2003).
Watanabe et al., J. Mol. Cel. Card. 37 (3) 691-698 (2004).
Weerasinghe et al., J. Virol. 65(10), 5531-3334 (1991).
Whitesell et al., Proc. Nat. Acad. Sci. 90: 4665-4669 (1993).
Wooddell, Christine I. et al., Long-Term RNA Interference from Optimized siRNA Expression Constructs in Adult Mice, Biochemical and Biophysical Research Communications, 2005, 334: pp. 117-127.
Xia et al., Nat. Biotech. 20, 1006-1010 (2002).
Xia et al., Nat. Med. 10(8) 816-820 (2004).
Yamamoto et al., Cell 101, 57-66 (2000).
Yamamoto, A. et al., "Reversal of Neuropathology and Motor Dysfunction in a Conditional Model of Huntington's Disease", Cell 101(1):pp. 57-66, 2000.
Yu et al., Proc. Nat. Acad. Sci. 90 6340-6344 (1993).
Yu et al., Proc. Nat. Acad. Sci. 99 6047-6052 (2002).
Zhang et al., "Global Non-Viral Gene Transfer to the Primate Brain Following Intravenous Administration," Molecular Therapy, Jan. 2003; 7(1): 11-18.
Zhang et al., "In vivo knockdown of gene expression in brain cancer with intravenous RNAi in adult rats," J. Gene Med., 2003; 5:1039-1045; published online Aug. 4, 2003.
Zhang et al., "Intravenous RNA Interference Gene Therapy Targeting the Human Epidermal Growth Factor Receptor Prolongs Survival in Intracranial Brain Cancer," Clinical Cancer Research, Jun. 1, 2004; 10:3667-3677.
Zhang et al., 1996 J. Mol. Neurosci. 7: 13-28 (1996).
Zhao et al., J. Biol. Chem. 271(49), 31407-31411 (Dec. 1996).
Zlokovic et al., Neurosurgery 40 805-813 (1997).

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